Publications
2023
Ponce José R Jaramillo, Théobald-Dietrich Anne, Bénas Philippe, Paulus Caroline, Sauter Claude, Frugier Magali
Solution x-ray scattering highlights discrepancies in Plasmodium multi-aminoacyl-tRNA synthetase complexes Article de journal
Dans: Protein Sci, p. e4564, 2023, ISSN: 1469-896X.
Résumé | Liens | BibTeX | Étiquettes: FRUGIER, SAUTER, Unité ARN
@article{pmid36606712,
title = {Solution x-ray scattering highlights discrepancies in Plasmodium multi-aminoacyl-tRNA synthetase complexes},
author = {José R Jaramillo Ponce and Anne Théobald-Dietrich and Philippe Bénas and Caroline Paulus and Claude Sauter and Magali Frugier},
doi = {10.1002/pro.4564},
issn = {1469-896X},
year = {2023},
date = {2023-01-01},
urldate = {2023-01-01},
journal = {Protein Sci},
pages = {e4564},
abstract = {tRip is a tRNA import protein specific to Plasmodium, the causative agent of malaria. In addition to its membrane localization and tRNA trafficking properties, tRip has the capacity to associate with three aminoacyl-tRNA synthetases (aaRS), the glutamyl- (ERS), glutaminyl- (QRS), and methionyl- (MRS) tRNA synthetases. In eukaryotes, such multi-aaRSs complexes (MSC) regulate the moonlighting activities of aaRSs. In Plasmodium, tRip and the three aaRSs all contain an N-terminal GST-like domain involved in the assembly of two independent complexes: the Q-complex (tRip:ERS:QRS) and the M-complex (tRip:ERS:MRS) with a 2:2:2 stoichiometry and in which the association of the GST-like domains of tRip and ERS (tRip-N:ERS-N) is central. In this study, the crystal structure of the N-terminal GST-like domain of ERS was solved and made possible further investigation of the solution architecture of the Q- and M-complexes by small-angle X-ray scattering (SAXS). This strategy relied on the engineering of a tRip-N-ERS-N chimeric protein to study the structural scaffold of both Plasmodium MSCs and confirm the unique homodimerization pattern of tRip in solution. The biological impact of these structural arrangements is discussed. This article is protected by copyright. All rights reserved.},
keywords = {FRUGIER, SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
2021
de Wijn R, Rollet K, Olieric V, Hennig O, Thome N, Nous C, Paulus C, Lorber B, Betat H, Morl M, Sauter C
Crystallization and Structural Determination of an Enzyme:Substrate Complex by Serial Crystallography in a Versatile Microfluidic Chip Article de journal
Dans: J Vis Exp, no. 169, 2021, ISBN: 33818565, (1940-087X (Electronic) 1940-087X (Linking) Journal Article Video-Audio Media).
Résumé | Liens | BibTeX | Étiquettes: SAUTER, Unité ARN
@article{deWijn2021,
title = {Crystallization and Structural Determination of an Enzyme:Substrate Complex by Serial Crystallography in a Versatile Microfluidic Chip},
author = {R de Wijn and K Rollet and V Olieric and O Hennig and N Thome and C Nous and C Paulus and B Lorber and H Betat and M Morl and C Sauter},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=33818565},
doi = {10.3791/61972},
isbn = {33818565},
year = {2021},
date = {2021-01-01},
journal = {J Vis Exp},
number = {169},
abstract = {The preparation of well diffracting crystals and their handling before their X-ray analysis are two critical steps of biocrystallographic studies. We describe a versatile microfluidic chip that enables the production of crystals by the efficient method of counter-diffusion. The convection-free environment provided by the microfluidic channels is ideal for crystal growth and useful to diffuse a substrate into the active site of the crystalline enzyme. Here we applied this approach to the CCA-adding enzyme of the psychrophilic bacterium Planococcus halocryophilus in the presented example. After crystallization and substrate diffusion/soaking, the crystal structure of the enzyme:substrate complex was determined at room temperature by serial crystallography and the analysis of multiple crystals directly inside the chip. The whole procedure preserves the genuine diffraction properties of the samples because it requires no crystal handling.},
note = {1940-087X (Electronic)
1940-087X (Linking)
Journal Article
Video-Audio Media},
keywords = {SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Wijn R. De, Rollet K., G.M.Ernst F., Wellner K., Betat H., Mörl M., Sauter M.
CCA-addition in the cold: Structural characterization of the psychrophilic CCA-adding enzyme from the permafrost bacterium Planococcus halocryophilus Article de journal
Dans: Computational and Structural Biotechnology Journal, vol. 19, p. 5845-5855, 2021, ISBN: ISBN/2001-0370.
Résumé | Liens | BibTeX | Étiquettes: CCA-adding enzyme, Cold adaptation, Psychrophilic protein, Psychrophilic RNA polymerase, SAUTER, SAXS, tRNA, Unité ARN, X-ray crystallography
@article{nokey,
title = {CCA-addition in the cold: Structural characterization of the psychrophilic CCA-adding enzyme from the permafrost bacterium Planococcus halocryophilus},
author = {R. De Wijn and K. Rollet and F. G.M.Ernst and K. Wellner and H. Betat and M. Mörl and M. Sauter},
url = {https://www.sciencedirect.com/science/article/pii/S2001037021004402?via%3Dihub},
doi = {10.1016/j.csbj.2021.10.018},
isbn = {ISBN/2001-0370},
year = {2021},
date = {2021-01-01},
journal = {Computational and Structural Biotechnology Journal},
volume = {19},
pages = {5845-5855},
abstract = {CCA-adding enzymes are highly specific RNA polymerases that add and maintain the sequence C-C-A at tRNA 3ム-ends. Recently, we could reveal that cold adaptation of such a polymerase is not only achieved at the expense of enzyme stability, but also at the cost of polymerization fidelity. Enzymes from psychrophilic organisms usually show an increased structural flexibility to enable catalysis at low temperatures. Here, polymerases face a dilemma, as there is a discrepancy between the need for a tightly controlled flexibility during polymerization and an increased flexibility as strategy for cold adaptation. Based on structural and biochemical analyses, we contribute to clarify the cold adaptation strategy of the psychrophilic CCA-adding enzyme from Planococcus halocryophilus, a gram-positive bacterium thriving in the arctic permafrost at low temperatures down to −15 °C. A comparison with the closely related enzyme from the thermophilic bacterium Geobacillus stearothermophilus reveals several features of cold adaptation - a significantly reduced amount of alpha-helical elements in the C-terminal tRNA-binding region and a structural adaptation in one of the highly conserved catalytic core motifs located in the N-terminal catalytic core of the enzyme},
keywords = {CCA-adding enzyme, Cold adaptation, Psychrophilic protein, Psychrophilic RNA polymerase, SAUTER, SAXS, tRNA, Unité ARN, X-ray crystallography},
pubstate = {published},
tppubtype = {article}
}
2020
Hennig O, Philipp S, Bonin S, Rollet K, Kolberg T, Jühling T, Betat H, Sauter C, Mörl M
Adaptation of the Romanomermis culicivorax CCA-Adding Enzyme to Miniaturized Armless tRNA Substrates Article de journal
Dans: International Journal of Molecular Sciences, vol. 21, no. 23, p. E9047, 2020.
Résumé | Liens | BibTeX | Étiquettes: CCA-adding enzyme, co-evolution, evolutionary plasticity, minimalized armless tRNAs, SAUTER, tRNA nucleotidyltransferase, Unité ARN
@article{12020,
title = {Adaptation of the Romanomermis culicivorax CCA-Adding Enzyme to Miniaturized Armless tRNA Substrates },
author = {O Hennig and S Philipp and S Bonin and K Rollet and T Kolberg and T Jühling and H Betat and C Sauter and M Mörl
},
url = {https://www.mdpi.com/1422-0067/21/23/9047},
doi = {10.3390/ijms21239047 },
year = {2020},
date = {2020-11-28},
journal = {International Journal of Molecular Sciences},
volume = {21},
number = {23},
pages = {E9047},
abstract = {The mitochondrial genome of the nematode Romanomermis culicivorax encodes for miniaturized hairpin-like tRNA molecules that lack D- as well as T-arms, strongly deviating from the consensus cloverleaf. The single tRNA nucleotidyltransferase of this organism is fully active on armless tRNAs, while the human counterpart is not able to add a complete CCA-end. Transplanting single regions of the Romanomermis enzyme into the human counterpart, we identified a beta-turn element of the catalytic core that-when inserted into the human enzyme-confers full CCA-adding activity on armless tRNAs. This region, originally identified to position the 3'-end of the tRNA primer in the catalytic core, dramatically increases the enzyme's substrate affinity. While conventional tRNA substrates bind to the enzyme by interactions with the T-arm, this is not possible in the case of armless tRNAs, and the strong contribution of the beta-turn compensates for an otherwise too weak interaction required for the addition of a complete CCA-terminus. This compensation demonstrates the remarkable evolutionary plasticity of the catalytic core elements of this enzyme to adapt to unconventional tRNA substrates. },
keywords = {CCA-adding enzyme, co-evolution, evolutionary plasticity, minimalized armless tRNAs, SAUTER, tRNA nucleotidyltransferase, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Théobald-Dietrich A, de Wijn R, Rollet K, Bluhm A, Rudinger-Thirion J, Paulus C, Lorber B, Thureau A, Frugier M, Sauter C
Structural Analysis of RNA by Small-Angle X-ray Scattering Chapitre d'ouvrage
Dans: Arluison, V; Wien, F (Ed.): RNA Spectroscopy: Methods and Protocols, vol. 2113, p. 189-215, Springer Protocols, Humana Press, New York, NY, 2020, ISBN: 32006316.
Résumé | Liens | BibTeX | Étiquettes: FRUGIER, IRES Integrative structural biology RNA SEC-SAXS Structure tRNA, SAUTER, Unité ARN
@inbook{,
title = {Structural Analysis of RNA by Small-Angle X-ray Scattering},
author = {A Théobald-Dietrich and R de Wijn and K Rollet and A Bluhm and J Rudinger-Thirion and C Paulus and B Lorber and A Thureau and M Frugier and C Sauter},
editor = {V Arluison and F Wien},
url = {https://pubmed.ncbi.nlm.nih.gov/32006316},
doi = {10.1007/978-1-0716-0278-2_14},
isbn = {32006316},
year = {2020},
date = {2020-01-01},
booktitle = {RNA Spectroscopy: Methods and Protocols},
volume = {2113},
pages = {189-215},
publisher = {Springer Protocols, Humana Press},
address = {New York, NY},
series = {Methods in Molecular Biology},
abstract = {Over the past two decades small-angle X-ray scattering (SAXS) has become a popular method to characterize solutions of biomolecules including ribonucleic acid (RNA). In an integrative structural approach, SAXS is complementary to crystallography, NMR, and electron microscopy and provides information about RNA architecture and dynamics. This chapter highlights the practical advantages of combining size-exclusion chromatography and SAXS at synchrotron facilities. It is illustrated by practical case studies of samples ranging from single hairpins and tRNA to a large IRES. The emphasis is also put on sample preparation which is a critical step of SAXS analysis and on optimized protocols for in vitro RNA synthesis ensuring the production of mg amount of pure and homogeneous molecules.},
keywords = {FRUGIER, IRES Integrative structural biology RNA SEC-SAXS Structure tRNA, SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {inbook}
}
Orlov I, Hemmer C, Ackerer L, Lorber B, Ghannam A, Poignavent V, Hleibieh K, Sauter C, Schmitt-Keichinger C, Belval L, Hily J M, Marmonier A, Komar V, Gersch S, Schellenberger P, Bron P, Vigne E, Muyldermans S, Lemaire O, Demangeat G, Ritzenthaler C, Klaholz B P
Structural Basis of Nanobody Recognition of Grapevine Fanleaf Virus and of Virus Resistance Loss Article de journal
Dans: Proc Natl Acad Sci U S A, vol. 117, no. 20, p. 10848-10855, 2020, ISBN: 32371486.
Résumé | Liens | BibTeX | Étiquettes: GFLV nanobody structural biology virus, SAUTER, Unité ARN
@article{,
title = {Structural Basis of Nanobody Recognition of Grapevine Fanleaf Virus and of Virus Resistance Loss},
author = {I Orlov and C Hemmer and L Ackerer and B Lorber and A Ghannam and V Poignavent and K Hleibieh and C Sauter and C Schmitt-Keichinger and L Belval and J M Hily and A Marmonier and V Komar and S Gersch and P Schellenberger and P Bron and E Vigne and S Muyldermans and O Lemaire and G Demangeat and C Ritzenthaler and B P Klaholz},
url = {https://www.ncbi.nlm.nih.gov/pubmed/32371486?dopt=Abstract},
doi = {10.1073/pnas.1913681117},
isbn = {32371486},
year = {2020},
date = {2020-01-01},
journal = {Proc Natl Acad Sci U S A},
volume = {117},
number = {20},
pages = {10848-10855},
abstract = {Grapevine fanleaf virus (GFLV) is a picorna-like plant virus transmitted by nematodes that affects vineyards worldwide. Nanobody (Nb)-mediated resistance against GFLV has been created recently, and shown to be highly effective in plants, including grapevine, but the underlying mechanism is unknown. Here we present the high-resolution cryo electron microscopy structure of the GFLV-Nb23 complex, which provides the basis for molecular recognition by the Nb. The structure reveals a composite binding site bridging over three domains of one capsid protein (CP) monomer. The structure provides a precise mapping of the Nb23 epitope on the GFLV capsid in which the antigen loop is accommodated through an induced-fit mechanism. Moreover, we uncover and characterize several resistance-breaking GFLV isolates with amino acids mapping within this epitope, including C-terminal extensions of the CP, which would sterically interfere with Nb binding. Escape variants with such extended CP fail to be transmitted by nematodes linking Nb-mediated resistance to vector transmission. Together, these data provide insights into the molecular mechanism of Nb23-mediated recognition of GFLV and of virus resistance loss.},
keywords = {GFLV nanobody structural biology virus, SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
de Wijn R, Rollet K, Engilberge S, McEwen A G, Hennig O, Betat H, Mörl M, Riobé F, Maury O, Girard E, Bénas P, Lorber B, Sauter C
Monitoring the Production of High Diffraction-Quality Crystals of Two Enzymes in Real Time Using In Situ Dynamic Light Scattering Article de journal
Dans: Crystals, vol. 10, no. 2, p. 65, 2020.
Résumé | Liens | BibTeX | Étiquettes: enzyme crystallization dynamic light scattering nucleation nucleant Tb-Xo4 crystallophore microcrystals nanocrystals X-ray diffraction XtalController, SAUTER, Unité ARN
@article{,
title = {Monitoring the Production of High Diffraction-Quality Crystals of Two Enzymes in Real Time Using In Situ Dynamic Light Scattering},
author = {R de Wijn and K Rollet and S Engilberge and A G McEwen and O Hennig and H Betat and M Mörl and F Riobé and O Maury and E Girard and P Bénas and B Lorber and C Sauter},
url = {https://www.mdpi.com/2073-4352/10/2/65},
doi = {10.3390/cryst10020065},
year = {2020},
date = {2020-01-01},
journal = {Crystals},
volume = {10},
number = {2},
pages = {65},
abstract = {The reproducible preparation of well-diffracting crystals is a prerequisite for every structural study based on crystallography. An instrument called XtalController has recently been designed that allows the monitoring of crystallization assays using dynamic light scattering and microscopy, and integrates piezo pumps to alter the composition of the mother liquor during the experiment. We have applied this technology to study the crystallization of two enzymes, the CCA-adding enzyme of the psychrophilic bacterium Planococcus halocryophilus, and the lysozyme from hen egg white in the presence of a synthetic chemical nucleant. We were able to (i) detect early nucleation events and (ii) drive the crystallization system (through cycles of dissolution/crystallization) toward growth conditions yielding crystals with excellent diffraction properties. This technology opens a way to the rational production of samples for crystallography, ranging from nanocrystals for electron diffraction, microcrystals for serial or conventional X-ray diffraction, to larger crystals for neutron diffraction.},
keywords = {enzyme crystallization dynamic light scattering nucleation nucleant Tb-Xo4 crystallophore microcrystals nanocrystals X-ray diffraction XtalController, SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
2019
Salinas T, Farouk-Ameqrane S E, Ubrig E, Sauter C, Duchêne A M, Maréchal-Drouard L
Molecular basis for the differential interaction of plant mitochondrial VDAC proteins with tRNAs Article de journal
Dans: Nucleic Acids Res, vol. 47, no. 2, p. 1048-1049, 2019, ISBN: 30519697.
Liens | BibTeX | Étiquettes: SAUTER, Unité ARN
@article{,
title = {Molecular basis for the differential interaction of plant mitochondrial VDAC proteins with tRNAs},
author = {T Salinas and S E Farouk-Ameqrane and E Ubrig and C Sauter and A M Duchêne and L Maréchal-Drouard},
url = {https://www.ncbi.nlm.nih.gov/pubmed/30517754?dopt=Abstract},
doi = {10.1093/nar/gky1247},
isbn = {30519697},
year = {2019},
date = {2019-01-01},
journal = {Nucleic Acids Res},
volume = {47},
number = {2},
pages = {1048-1049},
keywords = {SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
de Wijn R, Hennig O, Roche J, Engilberge S, Rollet K, Fernandez-Millan P, Brillet K, Betat H, Mörl M, Roussel A, Girard E, Mueller-Dieckmann C, Fox G C, Olieric V, Gavira J A, Lorber B, Sauter C
A simple and versatile microfluidic device for efficient biomacromolecule crystallization and structural analysis by serial crystallography Article de journal
Dans: IUCrJ, vol. 6, no. Pt 3, p. 454-464, 2019, ISBN: 31098026.
Résumé | Liens | BibTeX | Étiquettes: ChipX3 counter-diffusion crystallization ligand soaking macromolecule microfluidics protein structure room temperature seeding serial crystallography trace fluorescent labeling, ENNIFAR, SAUTER, Unité ARN
@article{,
title = {A simple and versatile microfluidic device for efficient biomacromolecule crystallization and structural analysis by serial crystallography},
author = {R de Wijn and O Hennig and J Roche and S Engilberge and K Rollet and P Fernandez-Millan and K Brillet and H Betat and M Mörl and A Roussel and E Girard and C Mueller-Dieckmann and G C Fox and V Olieric and J A Gavira and B Lorber and C Sauter},
url = {https://www.ncbi.nlm.nih.gov/pubmed/31098026?dopt=Abstract},
doi = {10.1107/S2052252519003622},
isbn = {31098026},
year = {2019},
date = {2019-01-01},
journal = {IUCrJ},
volume = {6},
number = {Pt 3},
pages = {454-464},
abstract = {Determining optimal conditions for the production of well diffracting crystals is a key step in every biocrystallography project. Here, a microfluidic device is described that enables the production of crystals by counter-diffusion and their direct on-chip analysis by serial crystallography at room temperature. Nine 'non-model' and diverse biomacromolecules, including seven soluble proteins, a membrane protein and an RNA duplex, were crystallized and treated on-chip with a variety of standard techniques including micro-seeding, crystal soaking with ligands and crystal detection by fluorescence. Furthermore, the crystal structures of four proteins and an RNA were determined based on serial data collected on four synchrotron beamlines, demonstrating the general applicability of this multipurpose chip concept.},
keywords = {ChipX3 counter-diffusion crystallization ligand soaking macromolecule microfluidics protein structure room temperature seeding serial crystallography trace fluorescent labeling, ENNIFAR, SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
2018
Jühling T, Duchardt-Ferner E, Bonin S, Wöhnert J, Pütz J, Florentz C, Betat H, Sauter C, Mörl M
Small but large enough: structural properties of armless mitochondrial tRNAs from the nematode Romanomermis culicivorax Article de journal
Dans: Nucleic Acids Res, vol. 46, no. 17, p. 9170-9180, 2018, ISBN: 29986062.
Résumé | Liens | BibTeX | Étiquettes: FLORENTZ, SAUTER, Unité ARN
@article{,
title = {Small but large enough: structural properties of armless mitochondrial tRNAs from the nematode Romanomermis culicivorax},
author = {T Jühling and E Duchardt-Ferner and S Bonin and J Wöhnert and J Pütz and C Florentz and H Betat and C Sauter and M Mörl},
url = {https://www.ncbi.nlm.nih.gov/pubmed/29986062?dopt=Abstract},
doi = {10.1093/nar/gky593},
isbn = {29986062},
year = {2018},
date = {2018-01-01},
journal = {Nucleic Acids Res},
volume = {46},
number = {17},
pages = {9170-9180},
abstract = {As adapter molecules to convert the nucleic acid information into the amino acid sequence, tRNAs play a central role in protein synthesis. To fulfill this function in a reliable way, tRNAs exhibit highly conserved structural features common in all organisms and in all cellular compartments active in translation. However, in mitochondria of metazoans, certain dramatic deviations from the consensus tRNA structure are described, where some tRNAs lack the D- or T-arm without losing their function. In Enoplea, this miniaturization comes to an extreme, and functional mitochondrial tRNAs can lack both arms, leading to a considerable size reduction. Here, we investigate the secondary and tertiary structure of two such armless tRNAs from Romanomermis culicivorax. Despite their high AU content, the transcripts fold into a single and surprisingly stable hairpin structure, deviating from standard tRNAs. The three-dimensional form is boomerang-like and diverges from the standard L-shape. These results indicate that such unconventional miniaturized tRNAs can still fold into a tRNA-like shape, although their length and secondary structure are very unusual. They highlight the remarkable flexibility of the protein synthesis apparatus and suggest that the translational machinery of Enoplea mitochondria may show compensatory adaptations to accommodate these armless tRNAs for efficient translation.},
keywords = {FLORENTZ, SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
de Wijn R, Hennig O, Ernst F G M, Lorber B, Betat H, Mörl M, Sauter C
Combining crystallogenesis methods to produce diffraction-quality crystals of a psychrophilic tRNA-maturation enzyme Article de journal
Dans: Acta Crystallogr F Struct Biol Commun, vol. 74, no. Pt 11, p. 747-753, 2018, ISBN: 30387781.
Résumé | Liens | BibTeX | Étiquettes: CCA-adding enzyme Planococcus halocryophilus counter-diffusion crystallogenesis microseeding optimization tRNA maturation trace fluorescent labeling, SAUTER, Unité ARN
@article{,
title = {Combining crystallogenesis methods to produce diffraction-quality crystals of a psychrophilic tRNA-maturation enzyme},
author = {R de Wijn and O Hennig and F G M Ernst and B Lorber and H Betat and M Mörl and C Sauter},
url = {https://www.ncbi.nlm.nih.gov/pubmed/30387781?dopt=Abstract},
doi = {10.1107/S2053230X18014590},
isbn = {30387781},
year = {2018},
date = {2018-01-01},
journal = {Acta Crystallogr F Struct Biol Commun},
volume = {74},
number = {Pt 11},
pages = {747-753},
abstract = {The determination of conditions for the reproducible growth of well diffracting crystals is a critical step in every biocrystallographic study. On the occasion of a new structural biology project, several advanced crystallogenesis approaches were tested in order to increase the success rate of crystallization. These methods included screening by microseed matrix screening, optimization by counter-diffusion and crystal detection by trace fluorescent labeling, and are easily accessible to any laboratory. Their combination proved to be particularly efficient in the case of the target, a 48 kDa CCA-adding enzyme from the psychrophilic bacterium Planococcus halocryophilus. A workflow summarizes the overall strategy, which led to the production of crystals that diffracted to better than 2 Å resolution and may be of general interest for a variety of applications.},
keywords = {CCA-adding enzyme Planococcus halocryophilus counter-diffusion crystallogenesis microseeding optimization tRNA maturation trace fluorescent labeling, SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
2017
Schellenberger P, Sauter C, Lorber B, Bron P, Trapani S, Bergdoll M, Marmonier A, Schmitt-Kelchinger C, Lemaire O, Demangeat G, Ritzenthaler C
Correction: Structural Insights into Viral Determinants of Nematode Mediated Grapevine fanleaf virus Transmission. Article de journal
Dans: PLoS Pathog, vol. 13, no. 3, p. e1006268, 2017, ISBN: 28296962.
Résumé | Liens | BibTeX | Étiquettes: SAUTER, Unité ARN
@article{,
title = {Correction: Structural Insights into Viral Determinants of Nematode Mediated Grapevine fanleaf virus Transmission.},
author = {P Schellenberger and C Sauter and B Lorber and P Bron and S Trapani and M Bergdoll and A Marmonier and C Schmitt-Kelchinger and O Lemaire and G Demangeat and C Ritzenthaler},
url = {https://www.ncbi.nlm.nih.gov/pubmed/28296962?dopt=Abstract},
doi = {10.1371/journal.ppat.1006268},
isbn = {28296962},
year = {2017},
date = {2017-01-01},
journal = {PLoS Pathog},
volume = {13},
number = {3},
pages = {e1006268},
abstract = {[This corrects the article DOI: 10.1371/journal.ppat.1002034.].
Erratum for
Structural insights into viral determinants of nematode mediated Grapevine fanleaf virus transmission. [PLoS Pathog. 2011]},
keywords = {SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Erratum for
Structural insights into viral determinants of nematode mediated Grapevine fanleaf virus transmission. [PLoS Pathog. 2011]
Pinker F, Schelcher C, Fernández-Millán P, Gobert A, Birck C, Thureau A, Roblin P, Giege P, Sauter C
Biophysical analysis of Arabidopsis protein-only RNase P alone and in complex with tRNA provides a refined model of tRNA binding. Article de journal
Dans: J Biol Chem, vol. 292, no. 34, p. 13904-13913, 2017, ISBN: 28696260.
Résumé | Liens | BibTeX | Étiquettes: PRORP RNA processing X-ray crystallography pentatricopeptide repeat (PPR) precursor tRNA (pre-tRNA) ribonuclease P (RNase P) small-angle X-ray scattering (SAXS) tRNA maturation, SAUTER, Unité ARN
@article{,
title = {Biophysical analysis of Arabidopsis protein-only RNase P alone and in complex with tRNA provides a refined model of tRNA binding.},
author = {F Pinker and C Schelcher and P Fernández-Millán and A Gobert and C Birck and A Thureau and P Roblin and P Giege and C Sauter},
url = {https://www.ncbi.nlm.nih.gov/pubmed/28696260?dopt=Abstract},
doi = {10.1074/jbc.M117.782078},
isbn = {28696260},
year = {2017},
date = {2017-01-01},
journal = {J Biol Chem},
volume = {292},
number = {34},
pages = {13904-13913},
abstract = {RNase P is a universal enzyme that removes 5' leader sequences from tRNA precursors. The enzyme is therefore essential for maturation of functional tRNAs and mRNA translation. RNase P represents a unique example of an enzyme that can occur either as ribonucleoprotein or as protein alone. The latter form of the enzyme called PRORP (PRotein-Only RNase P) is widespread in eukaryotes, in which it can provide organellar or nuclear RNase P activities. Here, we have focused on Arabidopsis nuclear PRORP2 and its interaction with tRNA substrates. Affinity measurements helped assess the respective importance of individual pentatricopeptide repeat motifs in PRORP2 for RNA binding. We characterized the PRORP2 structure by X-ray crystallography and by small-angle X-ray scattering (SAXS) in solution, as well as that of its complex with a tRNA precursor by SAXS. Of note, our study reports the first structural data of a PRORP-tRNA complex. Combined with complementary biochemical and biophysical analyses, our structural data suggest that PRORP2 undergoes conformational changes to accommodate its substrate. In particular, the catalytic domain and the RNA binding domain can move around a central hinge. Altogether, this work provides a refined model of the PRORP-tRNA complex that illustrates how protein-only RNase P enzymes specifically bind tRNA and highlights the contribution of protein dynamics to achieve this specific interaction.},
keywords = {PRORP RNA processing X-ray crystallography pentatricopeptide repeat (PPR) precursor tRNA (pre-tRNA) ribonuclease P (RNase P) small-angle X-ray scattering (SAXS) tRNA maturation, SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
2016
Belval Lorène, Hemmer Caroline, Sauter Claude, Reinbold Catherine, Fauny Jean-Daniel, Berthold François, Ackerer Léa, Schmitt-Keichinger Corinne, Lemaire Olivier, Demangeat Gérard, Ritzenthaler Christophe
Display of whole proteins on inner and outer surfaces of Grapevine fanleaf virus-like particles Article de journal
Dans: Plant Biotechnology Journal, 2016, ISSN: 1467-7652.
Résumé | Liens | BibTeX | Étiquettes: I2CT, Imagerie, SAUTER
@article{belval_display_2016,
title = {Display of whole proteins on inner and outer surfaces of Grapevine fanleaf virus-like particles},
author = {Lorène Belval and Caroline Hemmer and Claude Sauter and Catherine Reinbold and Jean-Daniel Fauny and François Berthold and Léa Ackerer and Corinne Schmitt-Keichinger and Olivier Lemaire and Gérard Demangeat and Christophe Ritzenthaler},
doi = {10.1111/pbi.12582},
issn = {1467-7652},
year = {2016},
date = {2016-01-01},
journal = {Plant Biotechnology Journal},
abstract = {Virus-like particles (VLPs) derived from non-enveloped viruses result from the self-assembly of capsid proteins (CPs). They generally display similar structural features to viral particles but are non-infectious and their inner cavity and outer surface can potentially be adapted to serve as nanocarriers of great biotechnological interest. While a VLP outer surface is generally amenable to chemical or genetic modifications, encaging a cargo within particles can be more complex and is often limited to small molecules or peptides. Examples where both inner cavity and outer surface have been used to simultaneously encapsulate and expose entire proteins remain scarce. Here we describe the production of spherical VLPs exposing fluorescent proteins at either their outer surface or inner cavity as a result of the self-assembly of a single genetically modified viral structural protein, the CP of grapevine fanleaf virus (GFLV). We found that the N- and C-terminal ends of the GFLV CP allow the genetic fusion of proteins as large as 27 kDa and the plant-based production of nucleic-acid free VLPs. Remarkably, expression of N- or C-terminal CP fusions, resulted in the production of VLPs with recombinant proteins exposed either to the inner cavity or the outer surface, respectively, while coexpression of both fusion proteins led to the formation hybrid VLP, although rather inefficiently. Such properties are rather unique for a single viral structural protein and open new potential avenues for the design of safe and versatile nanocarriers, particularly for the targeted delivery of bioactive molecules. This article is protected by copyright. All rights reserved.},
keywords = {I2CT, Imagerie, SAUTER},
pubstate = {published},
tppubtype = {article}
}
Schelcher C, Sauter C, Giege P
Mechanistic and Structural Studies of Protein-Only RNase P Compared to Ribonucleoproteins Reveal the Two Faces of the Same Enzymatic Activity. Article de journal
Dans: Biomolecules, vol. 6, no. 3, p. 30, 2016, ISBN: 27348014.
Résumé | Liens | BibTeX | Étiquettes: PRORP RNase P crystal structures kinetic analyses tRNA biogenesis, SAUTER, Unité ARN
@article{,
title = {Mechanistic and Structural Studies of Protein-Only RNase P Compared to Ribonucleoproteins Reveal the Two Faces of the Same Enzymatic Activity.},
author = {C Schelcher and C Sauter and P Giege},
url = {http://www.ncbi.nlm.nih.gov/pubmed/27348014?dopt=Abstract},
doi = {10.3390/biom6030030},
isbn = {27348014},
year = {2016},
date = {2016-01-01},
journal = {Biomolecules},
volume = {6},
number = {3},
pages = {30},
abstract = {RNase P, the essential activity that performs the 5' maturation of tRNA precursors, can be achieved either by ribonucleoproteins containing a ribozyme present in the three domains of life or by protein-only enzymes called protein-only RNase P (PRORP) that occur in eukaryote nuclei and organelles. A fast growing list of studies has investigated three-dimensional structures and mode of action of PRORP proteins. Results suggest that similar to ribozymes, PRORP proteins have two main domains. A clear functional analogy can be drawn between the specificity domain of the RNase P ribozyme and PRORP pentatricopeptide repeat domain, and between the ribozyme catalytic domain and PRORP N4BP1, YacP-like Nuclease domain. Moreover, both types of enzymes appear to dock with the acceptor arm of tRNA precursors and make specific contacts with the corner of pre-tRNAs. While some clear differences can still be delineated between PRORP and ribonucleoprotein (RNP) RNase P, the two types of enzymes seem to use, fundamentally, the same catalytic mechanism involving two metal ions. The occurrence of PRORP and RNP RNase P represents a remarkable example of convergent evolution. It might be the unique witness of an ongoing replacement of catalytic RNAs by proteins for enzymatic activities.},
keywords = {PRORP RNase P crystal structures kinetic analyses tRNA biogenesis, SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Fernandez-Millan P, Schelcher C, Chihade J, Masquida B, Giege P, Sauter C
Transfer RNA: from pioneering crystallographic studies to contemporary tRNA biology. Article de journal
Dans: Arch Biochem Biophys, vol. 602, p. 95-105, 2016, ISBN: 26968773.
Résumé | Liens | BibTeX | Étiquettes: RNA:protein recognition crystallography genetic code protein synthesis transfer RNA translation, SAUTER, Unité ARN
@article{,
title = {Transfer RNA: from pioneering crystallographic studies to contemporary tRNA biology.},
author = {P Fernandez-Millan and C Schelcher and J Chihade and B Masquida and P Giege and C Sauter},
url = {http://www.ncbi.nlm.nih.gov/pubmed/26968773?dopt=Abstract},
doi = {10.1016/j.abb.2016.03.005},
isbn = {26968773},
year = {2016},
date = {2016-01-01},
journal = {Arch Biochem Biophys},
volume = {602},
pages = {95-105},
abstract = {Transfer RNAs (tRNAs) play a key role in protein synthesis as adaptor molecules between messenger RNA and protein sequences on the ribosome. Their discovery in the early sixties provoked a worldwide infatuation with the study of their architecture and their function in the decoding of genetic information. tRNAs are also emblematic molecules in crystallography: the determination of the first tRNA crystal structures represented a milestone in structural biology and tRNAs were for a long period the sole source of information on RNA folding, architecture, and post-transcriptional modifications. Crystallographic data on tRNAs in complex with aminoacyl-tRNA synthetases (aaRSs) also provided the first insight into protein:RNA interactions. Beyond the translation process and the history of structural investigations on tRNA, this review also illustrates the renewal of tRNA biology with the discovery of a growing number of tRNA partners in the cell, the involvement of tRNAs in a variety of regulatory and metabolic pathways, and emerging applications in biotechnology and synthetic biology.},
keywords = {RNA:protein recognition crystallography genetic code protein synthesis transfer RNA translation, SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Belval L, Hemmer C, Sauter C, Reinbold C, Fauny J D, Berthold F, Ackerer L, Schmitt-Keichinger C, Lemaire O, Demangeat G, Ritzenthaler C
Display of whole proteins on inner and outer surfaces of Grapevine fanleaf virus-like particles. Article de journal
Dans: Plant Biotechnol J, vol. 14, no. 12, p. 2288-2299, 2016, ISBN: 27178344.
Résumé | Liens | BibTeX | Étiquettes: SAUTER, Unité ARN
@article{,
title = {Display of whole proteins on inner and outer surfaces of Grapevine fanleaf virus-like particles.},
author = {L Belval and C Hemmer and C Sauter and C Reinbold and J D Fauny and F Berthold and L Ackerer and C Schmitt-Keichinger and O Lemaire and G Demangeat and C Ritzenthaler},
url = {http://www.ncbi.nlm.nih.gov/pubmed/27178344?dopt=Abstract},
doi = {10.1111/pbi.12582},
isbn = {27178344},
year = {2016},
date = {2016-01-01},
journal = {Plant Biotechnol J},
volume = {14},
number = {12},
pages = {2288-2299},
abstract = {Virus-like particles (VLPs) derived from non-enveloped viruses result from the self-assembly of capsid proteins (CPs). They generally display similar structural features to viral particles but are non-infectious and their inner cavity and outer surface can potentially be adapted to serve as nanocarriers of great biotechnological interest. While a VLP outer surface is generally amenable to chemical or genetic modifications, encaging a cargo within particles can be more complex and is often limited to small molecules or peptides. Examples where both inner cavity and outer surface have been used to simultaneously encapsulate and expose entire proteins remain scarce. Here we describe the production of spherical VLPs exposing fluorescent proteins at either their outer surface or inner cavity as a result of the self-assembly of a single genetically modified viral structural protein, the CP of grapevine fanleaf virus (GFLV). We found that the N- and C-terminal ends of the GFLV CP allow the genetic fusion of proteins as large as 27 kDa and the plant-based production of nucleic-acid free VLPs. Remarkably, expression of N- or C-terminal CP fusions, resulted in the production of VLPs with recombinant proteins exposed either to the inner cavity or the outer surface, respectively, while coexpression of both fusion proteins led to the formation hybrid VLP, although rather inefficiently. Such properties are rather unique for a single viral structural protein and open new potential avenues for the design of safe and versatile nanocarriers, particularly for the targeted delivery of bioactive molecules.},
keywords = {SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
2015
Sauter C, Lorber B, Gaudry A, Karim L, Schwenzer H, Wien F, Roblin P, Florentz C, Sissler M
Neurodegenerative disease-associated mutants of a human mitochondrial aminoacyl-tRNA synthetase present individual molecular signatures. Article de journal
Dans: Sci Rep, vol. 5, p. 17332, 2015, ISBN: 26620921.
Résumé | Liens | BibTeX | Étiquettes: SAUTER, Unité ARN
@article{,
title = {Neurodegenerative disease-associated mutants of a human mitochondrial aminoacyl-tRNA synthetase present individual molecular signatures.},
author = {C Sauter and B Lorber and A Gaudry and L Karim and H Schwenzer and F Wien and P Roblin and C Florentz and M Sissler},
url = {http://www.ncbi.nlm.nih.gov/pubmed/26620921?dopt=Abstract},
doi = {10.1038/srep17332},
isbn = {26620921},
year = {2015},
date = {2015-01-01},
journal = {Sci Rep},
volume = {5},
pages = {17332},
abstract = {Mutations in human mitochondrial aminoacyl-tRNA synthetases are associated with a variety of neurodegenerative disorders. The effects of these mutations on the structure and function of the enzymes remain to be established. Here, we investigate six mutants of the aspartyl-tRNA synthetase correlated with leukoencephalopathies. Our integrated strategy, combining an ensemble of biochemical and biophysical approaches, reveals that mutants are diversely affected with respect to their solubility in cellular extracts and stability in solution, but not in architecture. Mutations with mild effects on solubility occur in patients as allelic combinations whereas those with strong effects on solubility or on aminoacylation are necessarily associated with a partially functional allele. The fact that all mutations show individual molecular and cellular signatures and affect amino acids only conserved in mammals, points towards an alternative function besides aminoacylation.},
keywords = {SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Pinker F, Giege P, Sauter C
Crystallization and crystallographic analysis of an Arabidopsis nuclear proteinaceous RNase P Article de journal
Dans: Acta Crystallogr F Struct Biol Commun, vol. 71, no. Pt 11, p. 1372-1377, 2015, ISBN: 26527263.
Résumé | Liens | BibTeX | Étiquettes: RNase P nuclear PRORP PPR tRNA maturation enzyme, SAUTER, Unité ARN
@article{,
title = {Crystallization and crystallographic analysis of an Arabidopsis nuclear proteinaceous RNase P},
author = {F Pinker and P Giege and C Sauter},
url = {http://www.ncbi.nlm.nih.gov/pubmed/26527263?dopt=Abstract},
doi = {10.1107/S2053230X15017033},
isbn = {26527263},
year = {2015},
date = {2015-01-01},
journal = {Acta Crystallogr F Struct Biol Commun},
volume = {71},
number = {Pt 11},
pages = {1372-1377},
abstract = {RNase P activity is ubiquitous and involves the 5' maturation of precursor tRNAs. For a long time, it was thought that all RNases P were ribonucleoproteic enzymes. However, the characterization of RNase P in human mitochondria and in plants revealed a novel kind of RNase P composed of protein only, called PRORP for `proteinaceous RNase P'. Whereas in human mitochondria PRORP has two partners that are required for RNase P activity, PRORP proteins are active as single-subunit enzymes in plants. Three paralogues of PRORP are found in Arabidopsis thaliana. PRORP1 is responsible for RNase P in mitochondria and chloroplasts, while PRORP2 and PRORP3 are nuclear enzymes. Here, the purification and crystallization of the Arabidopsis PRORP2 protein are reported. Optimization of the initial crystallization conditions led to crystals that diffracted to 3 Å resolution.},
keywords = {RNase P nuclear PRORP PPR tRNA maturation enzyme, SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
2014
Sauter D
Plongée au cœur des molécules du vivant (Diving into the heart of the molecules of life) Divers
2014.
BibTeX | Étiquettes: SAUTER, Unité ARN
@misc{,
title = {Plongée au cœur des molécules du vivant (Diving into the heart of the molecules of life)},
author = {D Sauter},
year = {2014},
date = {2014-01-01},
keywords = {SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {misc}
}
Salinas T, Farouk-Ameqrane S El, Ubrig E, Sauter C, Duchêne A M, Maréchal-Drouard L
Molecular basis for the differential interaction of plant mitochondrial VDAC proteins with tRNAs. Article de journal
Dans: Nucleic Acids Res, vol. 42, no. 15, p. 9937-9948, 2014, ISBN: 25114051.
Résumé | Liens | BibTeX | Étiquettes: SAUTER, Unité ARN
@article{,
title = {Molecular basis for the differential interaction of plant mitochondrial VDAC proteins with tRNAs.},
author = {T Salinas and S El Farouk-Ameqrane and E Ubrig and C Sauter and A M Duchêne and L Maréchal-Drouard},
url = {http://www.ncbi.nlm.nih.gov/pubmed/25114051?dopt=Abstract},
doi = {10.1093/nar/gku728},
isbn = {25114051},
year = {2014},
date = {2014-01-01},
journal = {Nucleic Acids Res},
volume = {42},
number = {15},
pages = {9937-9948},
abstract = {In plants, the voltage-dependent anion-selective channel (VDAC) is a major component of a pathway involved in transfer RNA (tRNA) translocation through the mitochondrial outer membrane. However, the way in which VDAC proteins interact with tRNAs is still unknown. Potato mitochondria contain two major mitochondrial VDAC proteins, VDAC34 and VDAC36. These two proteins, composed of a N-terminal α-helix and of 19 β-strands forming a β-barrel structure, share 75% sequence identity. Here, using both northwestern and gel shift experiments, we report that these two proteins interact differentially with nucleic acids. VDAC34 binds more efficiently with tRNAs or other nucleic acids than VDAC36. To further identify specific features and critical amino acids required for tRNA binding, 21 VDAC34 mutants were constructed and analyzed by northwestern. This allowed us to show that the β-barrel structure of VDAC34 and the first 50 amino acids that contain the α-helix are essential for RNA binding. Altogether the work shows that during evolution, plant mitochondrial VDAC proteins have diverged so as to interact differentially with nucleic acids, and this may reflect their involvement in various specialized biological functions.},
keywords = {SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
2013
Neuenfeldt A, Lorber B, Ennifar E, Gaudry A, Sauter C, Sissler M, Florentz C
Thermodynamic properties distinguish human mitochondrial aspartyl-tRNA synthetase from bacterial homolog with same 3D architecture Article de journal
Dans: Nucleic Acids Research, vol. 41, no. 4, p. 2698-708, 2013.
Résumé | Liens | BibTeX | Étiquettes: ENNIFAR, FRUGIER, SAUTER, Unité ARN
@article{Neuenfeldt2013,
title = {Thermodynamic properties distinguish human mitochondrial aspartyl-tRNA synthetase from bacterial homolog with same 3D architecture},
author = {A Neuenfeldt and B Lorber and E Ennifar and A Gaudry and C Sauter and M Sissler and C Florentz
},
url = {https://pubmed.ncbi.nlm.nih.gov/23275545/},
doi = { 10.1093/nar/gks1322 },
year = {2013},
date = {2013-02-04},
journal = {Nucleic Acids Research},
volume = {41},
number = {4},
pages = {2698-708},
abstract = {In the mammalian mitochondrial translation apparatus, the proteins and their partner RNAs are coded by two genomes. The proteins are nuclear-encoded and resemble their homologs, whereas the RNAs coming from the rapidly evolving mitochondrial genome have lost critical structural information. This raises the question of molecular adaptation of these proteins to their peculiar partner RNAs. The crystal structure of the homodimeric bacterial-type human mitochondrial aspartyl-tRNA synthetase (DRS) confirmed a 3D architecture close to that of Escherichia coli DRS. However, the mitochondrial enzyme distinguishes by an enlarged catalytic groove, a more electropositive surface potential and an alternate interaction network at the subunits interface. It also presented a thermal stability reduced by as much as 12°C. Isothermal titration calorimetry analyses revealed that the affinity of the mitochondrial enzyme for cognate and non-cognate tRNAs is one order of magnitude higher, but with different enthalpy and entropy contributions. They further indicated that both enzymes bind an adenylate analog by a cooperative allosteric mechanism with different thermodynamic contributions. The larger flexibility of the mitochondrial synthetase with respect to the bacterial enzyme, in combination with a preserved architecture, may represent an evolutionary process, allowing nuclear-encoded proteins to cooperate with degenerated organelle RNAs. },
keywords = {ENNIFAR, FRUGIER, SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
2012
Giege R, Juhling F, Putz J, Stadler P, Sauter C, Florentz C
Structure of transfer RNAs: similarity and variability Article de journal
Dans: Wiley Interdiscip Rev RNA, vol. 3, no. 1, p. 37-61, 2012, ISBN: 21957054.
Résumé | Liens | BibTeX | Étiquettes: FLORENTZ, FRUGIER, SAUTER, Unité ARN
@article{,
title = {Structure of transfer RNAs: similarity and variability},
author = {R Giege and F Juhling and J Putz and P Stadler and C Sauter and C Florentz},
url = {http://www.ncbi.nlm.nih.gov/pubmed/21957054?dopt=Abstrac},
doi = {10.1002/wrna.103},
isbn = {21957054},
year = {2012},
date = {2012-01-01},
journal = {Wiley Interdiscip Rev RNA},
volume = {3},
number = {1},
pages = {37-61},
abstract = {Transfer RNAs (tRNAs) are ancient molecules whose origin goes back to the beginning of life on Earth. Key partners in the ribosome-translation machinery, tRNAs read genetic information on messenger RNA and deliver codon specified amino acids attached to their distal 3′-extremity for peptide bond synthesis on the ribosome. In addition to this universal function, tRNAs participate in a wealth of other biological processes and undergo intricate maturation events. Our understanding of tRNA biology has been mainly phenomenological, but ongoing progress in structural biology is giving a robust physico-chemical basis that explains many facets of tRNA functions. Advanced sequence analysis of tRNA genes and their RNA transcripts have uncovered rules that underly tRNA 2D folding and 3D L-shaped architecture, as well as provided clues about their evolution. The increasing number of X-ray structures of free, protein- and ribosome-bound tRNA, reveal structural details accounting for the identity of the 22 tRNA families (one for each proteinogenic amino acid) and for the multifunctionality of a given family. Importantly, the structural role of post-transcriptional tRNA modifications is being deciphered. On the other hand, the plasticity of tRNA structure during function has been illustrated using a variety of technical approaches that allow dynamical insights. The large range of structural properties not only allows tRNAs to be the key actors of translation, but also sustain a diversity of unrelated functions from which only a few have already been pinpointed. Many surprises can still be expected. WIREs RNA 2012, 3:3761. doi: 10.1002/wrna.103},
keywords = {FLORENTZ, FRUGIER, SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Gaudry A, Lorber B, Neuenfeldt A, Sauter C, Florentz C, Sissler M
Re-designed N-terminus enhances expression, solubility and crystallizability of mitochondrial protein. Article de journal
Dans: Protein Eng Des Sel, vol. 25, no. 9, p. 473-481, 2012, ISBN: 22871419.
Résumé | Liens | BibTeX | Étiquettes: FLORENTZ, FRUGIER, SAUTER, SISSLER, Unité ARN
@article{,
title = {Re-designed N-terminus enhances expression, solubility and crystallizability of mitochondrial protein.},
author = {A Gaudry and B Lorber and A Neuenfeldt and C Sauter and C Florentz and M Sissler},
url = {http://www.ncbi.nlm.nih.gov/pubmed/22871419?report=&dispmax=200&tool=PubCrawler_2.23},
isbn = {22871419},
year = {2012},
date = {2012-01-01},
journal = {Protein Eng Des Sel},
volume = {25},
number = {9},
pages = {473-481},
abstract = {Mitochondrial aminoacyl-tRNA synthetases are key enzymes in translation. They are encoded by the nuclear genome, synthesized as precursors in the cytosol and imported. Most are matured by cleavage of their N-terminal targeting sequence. The poor expression of mature proteins in prokaryotic systems, along with their low solubility and stability after purification are major obstacles for biophysical and crystallographic studies. The purpose of the present work was to analyze the influence of additives on a slightly soluble aspartyl-tRNA synthetase and of the N-terminal sequence of the protein on its expression and solubility. On the one hand, the solubility of the enzyme was augmented to some extent in the presence of a chemical analog of the intermediary product aspartyl-adenylate, 5'-O-[N-(L aspartyl) sulfamoyl] adenosine. On the other hand, expression was enhanced by extending the N-terminus by seven natural amino acids from the predicted targeting sequence. The re-designed enzyme was active, monodisperse, more soluble and yielded crystals that are suitable for structure determination. This result underlines the importance of the N-terminal residue sequence for solubility. It suggests that additional criteria should be taken into account for the prediction of cleavage sites in mitochondrial targeting sequences.},
keywords = {FLORENTZ, FRUGIER, SAUTER, SISSLER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
2011
Schellenberger P, Sauter C, Lorber B, Bron P, Trapani S, Bergdoll M, Marmonier A, Schmitt-Kelchinger C, Lemaire O, Demangeat G, Ritzenthaler C
Structural insights into viral determinants of nematode mediated Grapevine fanleaf virus transmission. Article de journal
Dans: PLoS Pathog, vol. 7, no. 5, p. e1002034, 2011, ISBN: 21625570.
Résumé | Liens | BibTeX | Étiquettes: FLORENTZ, FRUGIER, SAUTER, Unité ARN
@article{,
title = {Structural insights into viral determinants of nematode mediated Grapevine fanleaf virus transmission.},
author = {P Schellenberger and C Sauter and B Lorber and P Bron and S Trapani and M Bergdoll and A Marmonier and C Schmitt-Kelchinger and O Lemaire and G Demangeat and C Ritzenthaler},
url = {http://www.ncbi.nlm.nih.gov/pubmed/21625570},
doi = {10.1371/journal.ppat.1002034},
isbn = {21625570},
year = {2011},
date = {2011-01-01},
journal = {PLoS Pathog},
volume = {7},
number = {5},
pages = {e1002034},
abstract = {Many animal and plant viruses rely on vectors for their transmission from host to host. Grapevine fanleaf virus (GFLV), a picorna-like virus from plants, is transmitted specifically by the ectoparasitic nematode Xiphinema index. The icosahedral capsid of GFLV, which consists of 60 identical coat protein subunits (CP), carries the determinants of this specificity. Here, we provide novel insight into GFLV transmission by nematodes through a comparative structural and functional analysis of two GFLV variants. We isolated a mutant GFLV strain (GFLV-TD) poorly transmissible by nematodes, and showed that the transmission defect is due to a glycine to aspartate mutation at position 297 (Gly297Asp) in the CP. We next determined the crystal structures of the wild-type GFLV strain F13 at 3.0 Å and of GFLV-TD at 2.7 Å resolution. The Gly297Asp mutation mapped to an exposed loop at the outer surface of the capsid and did not affect the conformation of the assembled capsid, nor of individual CP molecules. The loop is part of a positively charged pocket that includes a previously identified determinant of transmission. We propose that this pocket is a ligand-binding site with essential function in GFLV transmission by X. index. Our data suggest that perturbation of the electrostatic landscape of this pocket affects the interaction of the virion with specific receptors of the nematode's feeding apparatus, and thereby severely diminishes its transmission efficiency. These data provide a first structural insight into the interactions between a plant virus and a nematode vector.},
keywords = {FLORENTZ, FRUGIER, SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Schellenberger P, Demangeat G, Lemaire O, Ritzenthaler C, Bergdoll M, Olieric V, Sauter C, Lorber B
Strategies for the crystallization of viruses: Using phase diagrams and gels to produce 3D crystals of Grapevine fanleaf virus Article de journal
Dans: J Struct Biol, vol. 174, no. 2, p. 344-351, 2011, ISSN: 1095-8657 (Electronic) 1047-8477 (Linking), (Schellenberger, Pascale Demangeat, Gerard Lemaire, Olivier Ritzenthaler, Christophe Bergdoll, Marc Olieric, Vincent Sauter, Claude Lorber, Bernard United States Journal of structural biology J Struct Biol. 2011 May;174(2):344-51. Epub 2011 Feb 23.).
Résumé | Liens | BibTeX | Étiquettes: FLORENTZ, FRUGIER, SAUTER, Unité ARN
@article{,
title = {Strategies for the crystallization of viruses: Using phase diagrams and gels to produce 3D crystals of Grapevine fanleaf virus},
author = {P Schellenberger and G Demangeat and O Lemaire and C Ritzenthaler and M Bergdoll and V Olieric and C Sauter and B Lorber},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=21352920},
doi = {10.1016/j.jsb.2011.02.007},
issn = {1095-8657 (Electronic) 1047-8477 (Linking)},
year = {2011},
date = {2011-01-01},
journal = {J Struct Biol},
volume = {174},
number = {2},
pages = {344-351},
abstract = {The small icosahedral plant RNA nepovirus Grapevine fanleaf virus (GFLV) is specifically transmitted by a nematode and causes major damage to vineyards worldwide. To elucidate the molecular mechanisms underlying the recognition between the surface of its protein capsid and cellular components of its vector, host and viral proteins synthesized upon infection, the wild type GFLV strain F13 and a natural mutant (GFLV-TD) carrying a Gly(297)Asp mutation were purified, characterized and crystallized. Subsequently, the geometry and volume of their crystals was optimized by establishing phase diagrams. GFLV-TD was twice as soluble as the parent virus in the crystallization solution and its crystals diffracted X-rays to a resolution of 2.7A. The diffraction limit of GFLV-F13 crystals was extended from 5.5 to 3A by growth in agarose gel. Preliminary crystallographic analyses indicate that both types of crystals are suitable for structure determination. Keys for the successful production of GFLV crystals include the rigorous quality control of virus preparations, crystal quality improvement using phase diagrams, and crystal lattice reinforcement by growth in agarose gel. These strategies are applicable to the production of well-diffracting crystals of other viruses and macromolecular assemblies.},
note = {Schellenberger, Pascale
Demangeat, Gerard
Lemaire, Olivier
Ritzenthaler, Christophe
Bergdoll, Marc
Olieric, Vincent
Sauter, Claude
Lorber, Bernard
United States
Journal of structural biology
J Struct Biol. 2011 May;174(2):344-51. Epub 2011 Feb 23.},
keywords = {FLORENTZ, FRUGIER, SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
2009
Sauter C, Balg C, Moreno A, Dhouib K, Théobald-Dietrich A, Chenevert R, Giege R, Lorber B
Agarose gel facilitates enzyme crystal soaking with a ligand analog. Article de journal
Dans: J Appl Cryst, vol. 42, p. 279-283, 2009, ISBN: 10.1107/S0021889809003446.
Résumé | Liens | BibTeX | Étiquettes: Crystallization Agarose gel Ligands Inhibitors Soaking, FRUGIER, SAUTER, Unité ARN
@article{,
title = {Agarose gel facilitates enzyme crystal soaking with a ligand analog.},
author = {C Sauter and C Balg and A Moreno and K Dhouib and A Théobald-Dietrich and R Chenevert and R Giege and B Lorber},
url = {http://scripts.iucr.org/cgi-bin/paper?he5431},
isbn = {10.1107/S0021889809003446},
year = {2009},
date = {2009-01-01},
journal = {J Appl Cryst},
volume = {42},
pages = {279-283},
abstract = {Orthorhombic crystals of the enzyme aspartyl-tRNA synthetase (AspRS) were prepared in agarose gel, a chemical alternative to microgravity or nano-volume drops. Besides providing a convection-free medium, the network of the polysaccharide improved the stability of the crystalline lattice during soaking with L-aspartol adenylate, a synthetic and non-hydrolysable analog of the catalytic intermediate aspartyl adenylate. When crystals were embedded in the polysaccharide matrix the ligand reached their surfaces more uniformly. Gel-grown crystals exhibited well defined reflections even at high resolution and low mosaicity values, despite their fairly high solvent content and the relatively harsh flash cooling procedure. By contrast, soaked AspRS crystals prepared in solution broke apart within 10-30 s after the ligand was introduced into the mother liquor, and subsequently these fragments became an amorphous precipitate. A general objection to the use of gels in protein crystallization is that chemical interactions may occur between the polysaccharide matrix and proteins or ligands. The example of AspRS shows that this is not a major concern. This method may be generally applicable to crystal soaking with other small molecules or heavy atoms.},
keywords = {Crystallization Agarose gel Ligands Inhibitors Soaking, FRUGIER, SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Messmer M, Putz J, Suzuki T, Sauter C, Sissler M, Florentz C
Tertiary network in mammalian mitochondrial tRNAAsp revealed by solution probing and phylogeny Article de journal
Dans: Nucleic Acids Res, vol. 37, no. 20, p. 6881-6895, 2009, ISBN: 19767615, (1362-4962 (Electronic) 0305-1048 (Linking) Journal Article Research Support, Non-U.S. Gov't).
Résumé | Liens | BibTeX | Étiquettes: Asp/*chemistry/*metabolism Transcription, Base Sequence Databases, FLORENTZ, FRUGIER, Genetic, Nucleic Acid Humans Molecular Sequence Data Nucleic Acid Conformation Phylogeny RNA/*chemistry/*metabolism RNA, SAUTER, SISSLER, Transfer, Unité ARN
@article{,
title = {Tertiary network in mammalian mitochondrial tRNAAsp revealed by solution probing and phylogeny},
author = {M Messmer and J Putz and T Suzuki and C Sauter and M Sissler and C Florentz},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19767615},
isbn = {19767615},
year = {2009},
date = {2009-01-01},
journal = {Nucleic Acids Res},
volume = {37},
number = {20},
pages = {6881-6895},
abstract = {Primary and secondary structures of mammalian mitochondrial (mt) tRNAs are divergent from canonical tRNA structures due to highly skewed nucleotide content and large size variability of D- and T-loops. The nonconservation of nucleotides involved in the expected network of tertiary interactions calls into question the rules governing a functional L-shaped three-dimensional (3D) structure. Here, we report the solution structure of human mt-tRNA(Asp) in its native post-transcriptionally modified form and as an in vitro transcript. Probing performed with nuclease S1, ribonuclease V1, dimethylsulfate, diethylpyrocarbonate and lead, revealed several secondary structures for the in vitro transcribed mt-tRNA(Asp) including predominantly the cloverleaf. On the contrary, the native tRNA(Asp) folds into a single cloverleaf structure, highlighting the contribution of the four newly identified post-transcriptional modifications to correct folding. Reactivities of nucleotides and phosphodiester bonds in the native tRNA favor existence of a full set of six classical tertiary interactions between the D-domain and the variable region, forming the core of the 3D structure. Reactivities of D- and T-loop nucleotides support an absence of interactions between these domains. According to multiple sequence alignments and search for conservation of Leontis-Westhof interactions, the tertiary network core building rules apply to all tRNA(Asp) from mammalian mitochondria.},
note = {1362-4962 (Electronic)
0305-1048 (Linking)
Journal Article
Research Support, Non-U.S. Gov't},
keywords = {Asp/*chemistry/*metabolism Transcription, Base Sequence Databases, FLORENTZ, FRUGIER, Genetic, Nucleic Acid Humans Molecular Sequence Data Nucleic Acid Conformation Phylogeny RNA/*chemistry/*metabolism RNA, SAUTER, SISSLER, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Messmer M, Blais S P, Balg C, Chenevert R, Grenier L, Lague P, Sauter C, Sissler M, Giege R, Lapointe J, Florentz C
Peculiar inhibition of human mitochondrial aspartyl-tRNA synthetase by adenylate analogs Article de journal
Dans: Biochimie, vol. 91, no. 5, p. 596-603, 2009, ISBN: 19254750, (1638-6183 (Electronic) Journal Article Research Support, Non-U.S. Gov't).
Résumé | Liens | BibTeX | Étiquettes: Adenosine Monophosphate/*analogs & derivatives/*pharmacology Animals Aspartate-tRNA Ligase/*antagonists & inhibitors/*chemistry/metabolism Catalytic Domain Cattle Humans Mitochondria/*drug effects/*enzymology Molecular Structure Structure-Activity Relationship, FLORENTZ, SAUTER, SISSLER, Unité ARN
@article{,
title = {Peculiar inhibition of human mitochondrial aspartyl-tRNA synthetase by adenylate analogs},
author = {M Messmer and S P Blais and C Balg and R Chenevert and L Grenier and P Lague and C Sauter and M Sissler and R Giege and J Lapointe and C Florentz},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19254750},
isbn = {19254750},
year = {2009},
date = {2009-01-01},
journal = {Biochimie},
volume = {91},
number = {5},
pages = {596-603},
abstract = {Human mitochondrial aminoacyl-tRNA synthetases (mt-aaRSs), the enzymes which esterify tRNAs with the cognate specific amino acid, form mainly a different set of proteins than those involved in the cytosolic translation machinery. Many of the mt-aaRSs are of bacterial-type in regard of sequence and modular structural organization. However, the few enzymes investigated so far do have peculiar biochemical and enzymological properties such as decreased solubility, decreased specific activity and enlarged spectra of substrate tRNAs (of same specificity but from various organisms and kingdoms), as compared to bacterial aaRSs. Here the sensitivity of human mitochondrial aspartyl-tRNA synthetase (AspRS) to small substrate analogs (non-hydrolysable adenylates) known as inhibitors of Escherichia coli and Pseudomonas aeruginosa AspRSs is evaluated and compared to the sensitivity of eukaryal cytosolic human and bovine AspRSs. L-aspartol-adenylate (aspartol-AMP) is a competitive inhibitor of aspartylation by mitochondrial as well as cytosolic mammalian AspRSs, with K(i) values in the micromolar range (4-27 microM for human mt- and mammalian cyt-AspRSs). 5'-O-[N-(L-aspartyl)sulfamoyl]adenosine (Asp-AMS) is a 500-fold stronger competitive inhibitor of the mitochondrial enzyme than aspartol-AMP (10nM) and a 35-fold lower competitor of human and bovine cyt-AspRSs (300 nM). The higher sensitivity of human mt-AspRS for both inhibitors as compared to either bacterial or mammalian cytosolic enzymes, is not correlated with clear-cut structural features in the catalytic site as deduced from docking experiments, but may result from dynamic events. In the scope of new antibacterial strategies directed against aaRSs, possible side effects of such drugs on the mitochondrial human aaRSs should thus be considered.},
note = {1638-6183 (Electronic)
Journal Article
Research Support, Non-U.S. Gov't},
keywords = {Adenosine Monophosphate/*analogs & derivatives/*pharmacology Animals Aspartate-tRNA Ligase/*antagonists & inhibitors/*chemistry/metabolism Catalytic Domain Cattle Humans Mitochondria/*drug effects/*enzymology Molecular Structure Structure-Activity Relationship, FLORENTZ, SAUTER, SISSLER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Lorber B, Sauter C, Théobald-Dietrich A, Moreno A, Schellenberger P, Robert M C, Capelle B, Sanglier S, Potier N, Giege R
Crystal growth of proteins, nucleic acids, and viruses in gels Article de journal
Dans: Prog Biophys Mol Biol, vol. 101, no. 1-3, p. 13-25, 2009, ISBN: 20005247, (1873-1732 (Electronic) 0079-6107 (Linking) Journal article).
Résumé | Liens | BibTeX | Étiquettes: FRUGIER, SAUTER, Unité ARN
@article{,
title = {Crystal growth of proteins, nucleic acids, and viruses in gels},
author = {B Lorber and C Sauter and A Théobald-Dietrich and A Moreno and P Schellenberger and M C Robert and B Capelle and S Sanglier and N Potier and R Giege},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=20005247},
isbn = {20005247},
year = {2009},
date = {2009-01-01},
journal = {Prog Biophys Mol Biol},
volume = {101},
number = {1-3},
pages = {13-25},
abstract = {Medium-sized single crystals with perfect habits and no defect producing intense and well-resolved diffraction patterns are the dream of every protein crystallographer. Crystals of biological macromolecules possessing these characteristics can be prepared within a medium in which mass transport is restricted to diffusion. Chemical gels (like polysiloxane) and physical gels (such as agarose) provide such an environment and are therefore suitable for the crystallisation of biological macromolecules. Instructions for the preparation of each type of gel are given to urge crystal growers to apply diffusive media for enhancing crystallographic quality of their crystals. Examples of quality enhancement achieved with silica and agarose gels are given. Results obtained with other substances forming gel-like media (such as lipidic phases and cellulose derivatives) are presented. Finally, the use of gels in combination with capillary tubes for counter-diffusion experiments is discussed. Methods and techniques implemented with proteins can also be applied to nucleic acids and nucleoprotein assemblies such as viruses.},
note = {1873-1732 (Electronic)
0079-6107 (Linking)
Journal article},
keywords = {FRUGIER, SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Dhouib K, Malek C Khan, Pfleging W, Gauthier-Manuel B, Duffait R, Thuillier G, Ferrigno R, Jacquamet L, Ohana J, Ferrer J L, Théobald-Dietrich A, Giege R, Lorber B, Sauter C
Microfluidic chips for the crystallization of biomacromolecules by counter-diffusion and on-chip crystal X-ray analysis Article de journal
Dans: Lab Chip, vol. 9, no. 10, p. 1412-1421, 2009, ISBN: 19417908, (1473-0197 (Print) 1473-0189 (Linking) Journal Article Research Support, Non-U.S. Gov't).
Résumé | Liens | BibTeX | Étiquettes: FRUGIER, SAUTER, Unité ARN, X-Ray/*instrumentation Dimethylpolysiloxanes/chemistry Macromolecular Substances/*chemistry Microfluidic Analytical Techniques/*instrumentation Polymethyl Methacrylate/chemistry
@article{,
title = {Microfluidic chips for the crystallization of biomacromolecules by counter-diffusion and on-chip crystal X-ray analysis},
author = {K Dhouib and C Khan Malek and W Pfleging and B Gauthier-Manuel and R Duffait and G Thuillier and R Ferrigno and L Jacquamet and J Ohana and J L Ferrer and A Théobald-Dietrich and R Giege and B Lorber and C Sauter},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19417908},
isbn = {19417908},
year = {2009},
date = {2009-01-01},
journal = {Lab Chip},
volume = {9},
number = {10},
pages = {1412-1421},
abstract = {Microfluidic devices were designed to perform on micromoles of biological macromolecules and viruses the search and the optimization of crystallization conditions by counter-diffusion, as well as the on-chip analysis of crystals by X-ray diffraction. Chips composed of microchannels were fabricated in poly-dimethylsiloxane (PDMS), poly-methyl-methacrylate (PMMA) and cyclo-olefin-copolymer (COC) by three distinct methods, namely replica casting, laser ablation and hot embossing. The geometry of the channels was chosen to ensure that crystallization occurs in a convection-free environment. The transparency of the materials is compatible with crystal growth monitoring by optical microscopy. The quality of the protein 3D structures derived from on-chip crystal analysis by X-ray diffraction using a synchrotron radiation was used to identify the most appropriate polymers. Altogether the results demonstrate that for a novel biomolecule, all steps from the initial search of crystallization conditions to X-ray diffraction data collection for 3D structure determination can be performed in a single chip.},
note = {1473-0197 (Print)
1473-0189 (Linking)
Journal Article
Research Support, Non-U.S. Gov't},
keywords = {FRUGIER, SAUTER, Unité ARN, X-Ray/*instrumentation Dimethylpolysiloxanes/chemistry Macromolecular Substances/*chemistry Microfluidic Analytical Techniques/*instrumentation Polymethyl Methacrylate/chemistry},
pubstate = {published},
tppubtype = {article}
}
2008
Giege R, Touze E, Lorber B, Théobald-Dietrich A, Sauter C
Crystallogenesis trends of free and liganded aminoacyl-tRNA synthetases Article de journal
Dans: Crystal Growth & Design, vol. 8, no. 12, p. 4297-4306, 2008, ISBN: 10.1021/cg8007766.
Résumé | Liens | BibTeX | Étiquettes: FRUGIER, SAUTER, Unité ARN
@article{,
title = {Crystallogenesis trends of free and liganded aminoacyl-tRNA synthetases},
author = {R Giege and E Touze and B Lorber and A Théobald-Dietrich and C Sauter},
url = {http://pubs.acs.org/doi/abs/10.1021/cg8007766?prevSearch=%255BContrib%253A%2BSauter%2Bc%255D&searchHistoryKey=},
isbn = {10.1021/cg8007766},
year = {2008},
date = {2008-01-01},
journal = {Crystal Growth & Design},
volume = {8},
number = {12},
pages = {4297-4306},
abstract = {Aminoacyl-tRNA synthetases (aaRSs) catalyze the attachment of amino acids on transfer RNAs (tRNAs) and thus enable the correct expression of the genetic code during protein synthesis. After a brief historical review of early aaRS crystallizations we present results of a survey of the crystallization conditions of 220 aaRSs (either free or bound to small ligands) and of 59 aaRS:tRNA complexes whose structures are hosted by the RCSB Protein Data Bank. These structures at resolutions between 4.5 and 1.2 Å are those of synthetases from the three kingdoms of life. Their phylogenic distribution is heterogeneous: most stem from bacteria and archaea and some from eukarya and organelles. Interesting correlations are found between biochemical, physical−chemical, and crystallographic parameters. They highlight common and more specific features of the crystallogenesis of free or liganded aaRSs. The effects of the chemical nature of the crystallant(s), of ligands and other additives, as well as of the presence of impurities and of the presence of flexible polypeptide domains on protein or protein/RNA crystallization are examined. Features that favor the growth of crystals diffracting X-rays at high resolution are discussed.},
keywords = {FRUGIER, SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Blaise M, Olieric V, Sauter C, Lorber B, Roy B, Karmakar S, Banerjee R, Becker H D, Kern D
Dans: J Mol Biol, vol. 381, no. 5, p. 1224-1237, 2008, ISBN: 18602926, (1089-8638 (Electronic) Journal Article Research Support, Non-U.S. Gov't).
Résumé | Liens | BibTeX | Étiquettes: Adenosine Triphosphate/metabolism Amino Acid Sequence Amino Acyl-tRNA Synthetases/*chemistry Anticodon/*metabolism Base Sequence Binding Sites Catalysis Conserved Sequence Crystallography, Asp/*chemistry/genetics Regulatory Sequences, FFRUGIER, Molecular Molecular Sequence Data Nucleic Acid Conformation Nucleoside Q/*chemistry Protein Structure, Ribonucleic Acid/*genetics Thermus thermophilus/enzymology, SAUTER, Secondary RNA, Transfer, Unité ARN, X-Ray Escherichia coli/*enzymology Escherichia coli Proteins/*chemistry Glutamic Acid/*chemistry Models
@article{,
title = {Crystal structure of glutamyl-queuosine tRNAAsp synthetase complexed with L-glutamate: structural elements mediating tRNA-independent activation of glutamate and glutamylation of tRNAAsp anticodon},
author = {M Blaise and V Olieric and C Sauter and B Lorber and B Roy and S Karmakar and R Banerjee and H D Becker and D Kern},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18602926},
isbn = {18602926},
year = {2008},
date = {2008-01-01},
journal = {J Mol Biol},
volume = {381},
number = {5},
pages = {1224-1237},
abstract = {Glutamyl-queuosine tRNA(Asp) synthetase (Glu-Q-RS) from Escherichia coli is a paralog of the catalytic core of glutamyl-tRNA synthetase (GluRS) that catalyzes glutamylation of queuosine in the wobble position of tRNA(Asp). Despite important structural similarities, Glu-Q-RS and GluRS diverge strongly by their functional properties. The only feature common to both enzymes consists in the activation of Glu to form Glu-AMP, the intermediate of transfer RNA (tRNA) aminoacylation. However, both enzymes differ by the mechanism of selection of the cognate amino acid and by the mechanism of its activation. Whereas GluRS selects l-Glu and activates it only in the presence of the cognate tRNA(Glu), Glu-Q-RS forms Glu-AMP in the absence of tRNA. Moreover, while GluRS transfers the activated Glu to the 3' accepting end of the cognate tRNA(Glu), Glu-Q-RS transfers the activated Glu to Q34 located in the anticodon loop of the noncognate tRNA(Asp). In order to gain insight into the structural elements leading to distinct mechanisms of amino acid activation, we solved the three-dimensional structure of Glu-Q-RS complexed to Glu and compared it to the structure of the GluRS.Glu complex. Comparison of the catalytic site of Glu-Q-RS with that of GluRS, combined with binding experiments of amino acids, shows that a restricted number of residues determine distinct catalytic properties of amino acid recognition and activation by the two enzymes. Furthermore, to explore the structural basis of the distinct aminoacylation properties of the two enzymes and to understand why Glu-Q-RS glutamylates only tRNA(Asp) among the tRNAs possessing queuosine in position 34, we performed a tRNA mutational analysis to search for the elements of tRNA(Asp) that determine recognition by Glu-Q-RS. The analyses made on tRNA(Asp) and tRNA(Asn) show that the presence of a C in position 38 is crucial for glutamylation of Q34. The results are discussed in the context of the evolution and adaptation of the tRNA glutamylation system.},
note = {1089-8638 (Electronic)
Journal Article
Research Support, Non-U.S. Gov't},
keywords = {Adenosine Triphosphate/metabolism Amino Acid Sequence Amino Acyl-tRNA Synthetases/*chemistry Anticodon/*metabolism Base Sequence Binding Sites Catalysis Conserved Sequence Crystallography, Asp/*chemistry/genetics Regulatory Sequences, FFRUGIER, Molecular Molecular Sequence Data Nucleic Acid Conformation Nucleoside Q/*chemistry Protein Structure, Ribonucleic Acid/*genetics Thermus thermophilus/enzymology, SAUTER, Secondary RNA, Transfer, Unité ARN, X-Ray Escherichia coli/*enzymology Escherichia coli Proteins/*chemistry Glutamic Acid/*chemistry Models},
pubstate = {published},
tppubtype = {article}
}
2007
Touze E, Lorber B, Deniziak M, Becker H D, Kern D, Giege R, Sauter C
Disorder Can Exist inside Well-Diffracting Crystals Article de journal
Dans: Crystal Growth & Design, vol. 7, no. 11, p. 2195-2197, 2007, (DOI: 10.1021/cg7007057).
Résumé | Liens | BibTeX | Étiquettes: FRUGIER, GIEGE KERN FLORENTZ, SAUTER, Unité ARN
@article{,
title = {Disorder Can Exist inside Well-Diffracting Crystals},
author = {E Touze and B Lorber and M Deniziak and H D Becker and D Kern and R Giege and C Sauter},
url = {http://pubs.acs.org/doi/abs/10.1021/cg7007057?prevSearch=Touz%25C3%25A9&searchHistoryKey=},
year = {2007},
date = {2007-01-01},
journal = {Crystal Growth & Design},
volume = {7},
number = {11},
pages = {2195-2197},
abstract = {Unlike other glutaminyl-tRNA synthetases, the one from radioresistant bacterium Deinococcus radiodurans (Dr-GlnRS) possesses an additional C-terminal extension of 220 residues that shares some homology with the subunit of another enzyme of the translation machinery. Dr-GlnRS has been crystallized in an orthorhombic space group. The crystals diffract X-rays to a resolution of 2 Å. The determination of the structure of this atypical GlnRS showed that its N- and C-terminal appendices, which encompass in total one third of the proteinメs 852 amino acids, are actually disordered in the crystal lattice. This example demonstrates that macromolecule crystallization can tolerate large flexible regions in the solvent channels as long as they do not interfere with the packing contacts. This intriguing case is analyzed and discussed in light of current crystallogenesis strategies.},
note = {DOI: 10.1021/cg7007057},
keywords = {FRUGIER, GIEGE KERN FLORENTZ, SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Sauter C, Dhouib K, Lorber B
From macrofluidies to microfluidies for the crystallization of biological macromolecules Article de journal
Dans: Crystal Growth & Design, vol. 7, no. 11, p. 2247-2250, 2007, (DOI: 10.1021/cg700955f).
Résumé | Liens | BibTeX | Étiquettes: FRUGIER, GIEGE FLORENTZ, SAUTER, Unité ARN
@article{,
title = {From macrofluidies to microfluidies for the crystallization of biological macromolecules},
author = {C Sauter and K Dhouib and B Lorber},
url = {http://pubs.acs.org/doi/abs/10.1021/cg700955f?prevSearch=sauter&searchHistoryKey=},
year = {2007},
date = {2007-01-01},
journal = {Crystal Growth & Design},
volume = {7},
number = {11},
pages = {2247-2250},
abstract = {This review presents the goals and principles of microfluidic technologies applied to the crystallization of biological macromolecules. A comparison of the devices that are available commercially or described in the literature summarizes the current state-of-the-art in microfluidics. A novel chip based on the counter-diffusion of solute molecules playing the role of crystallization agents is described. Inside the microfluidic channels composing the chip mass transport essentially occurs by diffusion. The chip is made of a rigid polymer that is impermeable to gases and compatible with crystal examination and monitoring in polarized light. The selected material is also transparent to X-rays, and three-dimensional protein structures can be determined from crystals contained inside this device using X-ray diffraction data collected on a synchrotron source. The outstanding quality of the electron-density maps demonstrates that on-chip crystal analysis is feasible. The replacement of conventional crystallization setups by inexpensive microfluidic chips for screening best crystallization agents and automated crystal diffraction analysis is discussed.},
note = {DOI: 10.1021/cg700955f},
keywords = {FRUGIER, GIEGE FLORENTZ, SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
2006
Sauter C, Lorber B, Théobald-Dietrich A, Giege R, Khan-Malek C, Gauthier-Manuel B, Thuillier G, Ferrigno R
Dispositif microfluidique pour la cristallisation et l'analyse cristallographique de molécules. Divers
2006, ISBN: EP 2040809 A2.
BibTeX | Étiquettes: FRUGIER, GIEGE, SAUTER, Unité ARN
@misc{,
title = {Dispositif microfluidique pour la cristallisation et l'analyse cristallographique de molécules.},
author = {C Sauter and B Lorber and A Théobald-Dietrich and R Giege and C Khan-Malek and B Gauthier-Manuel and G Thuillier and R Ferrigno},
isbn = {EP 2040809 A2},
year = {2006},
date = {2006-01-01},
keywords = {FRUGIER, GIEGE, SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {misc}
}
Fender A, Sauter C, Messmer M, Putz J, Giege R, Florentz C, Sissler M
Loss of a primordial identity element for a mammalian mitochondrial aminoacylation system Article de journal
Dans: J Biol Chem, vol. 281, no. 23, p. 15980-15986, 2006, ISBN: 16597625, (0021-9258 (Print) Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Acylation Base Sequence Humans Kinetics Mutagenesis Nucleic Acid Conformation Plasmids RNA | Non-U.S. Gov't, Asp/chemistry/genetics/*metabolism Research Support, FLORENTZ, FRUGIER, Non-U.S. Gov't, SAUTER, SISSLER, Transfer, Unité ARN
@article{,
title = {Loss of a primordial identity element for a mammalian mitochondrial aminoacylation system},
author = {A Fender and C Sauter and M Messmer and J Putz and R Giege and C Florentz and M Sissler},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16597625},
isbn = {16597625},
year = {2006},
date = {2006-01-01},
journal = {J Biol Chem},
volume = {281},
number = {23},
pages = {15980-15986},
abstract = {In mammalian mitochondria the translational machinery is of dual origin with tRNAs encoded by a simplified and rapidly evolving mitochondrial (mt) genome and aminoacyl-tRNA synthetases (aaRS) coded by the nuclear genome, and imported. Mt-tRNAs are atypical with biased sequences, size variations in loops and stems, and absence of residues forming classical tertiary interactions, whereas synthetases appear typical. This raises questions about identity elements in mt-tRNAs and adaptation of their cognate mt-aaRSs. We have explored here the human mt-aspartate system in which a prokaryotic-type AspRS, highly similar to the Escherichia coli enzyme, recognizes a bizarre tRNA(Asp). Analysis of human mt-tRNA(Asp) transcripts confirms the identity role of the GUC anticodon as in other aspartylation systems but reveals the non-involvement of position 73. This position is otherwise known as the site of a universally conserved major aspartate identity element, G73, also known as a primordial identity signal. In mt-tRNA(Asp), position 73 can be occupied by any of the four nucleotides without affecting aspartylation. Sequence alignments of various AspRSs allowed placing Gly-269 at a position occupied by Asp-220, the residue contacting G73 in the crystallographic structure of E. coli AspRS-tRNA(Asp) complex. Replacing this glycine by an aspartate renders human mt-AspRS more discriminative to G73. Restriction in the aspartylation identity set, driven by a rapid mutagenic rate of the mt-genome, suggests a reverse evolution of the mt-tRNA(Asp) identity elements in regard to its bacterial ancestor.},
note = {0021-9258 (Print)
Journal Article},
keywords = {Acylation Base Sequence Humans Kinetics Mutagenesis Nucleic Acid Conformation Plasmids RNA | Non-U.S. Gov't, Asp/chemistry/genetics/*metabolism Research Support, FLORENTZ, FRUGIER, Non-U.S. Gov't, SAUTER, SISSLER, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
2005
Vergara A, Lorber B, Sauter C, Giege R, Zagari A
Lessons from crystals grown in the Advanced Protein Crystallisation Facility for conventional crystallisation applied to structural biology Article de journal
Dans: Biophys Chem, vol. 118, no. 2-3, p. 102-112, 2005, ISBN: 16150532, (0301-4622 (Print) Journal Article Review).
Résumé | Liens | BibTeX | Étiquettes: FRUGIER, GIEGE Animals *Biological Sciences Crystallization Equipment Design Humans Protein Conformation Proteins/*chemistry Research Support, Non-U.S. Gov't X-Ray Diffraction, SAUTER, Unité ARN
@article{,
title = {Lessons from crystals grown in the Advanced Protein Crystallisation Facility for conventional crystallisation applied to structural biology},
author = {A Vergara and B Lorber and C Sauter and R Giege and A Zagari},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16150532},
isbn = {16150532},
year = {2005},
date = {2005-01-01},
journal = {Biophys Chem},
volume = {118},
number = {2-3},
pages = {102-112},
abstract = {The crystallographic quality of protein crystals that were grown in microgravity has been compared to that of crystals that were grown in parallel on earth gravity under otherwise identical conditions. A goal of this comparison was to assess if a more accurate 3D-structure can be derived from crystallographic analysis of the former crystals. Therefore, the properties of crystals prepared with the Advanced Protein Crystallisation Facility (APCF) on earth and in orbit during the last decade were evaluated. A statistical analysis reveals that about half of the crystals produced under microgravity had a superior X-ray diffraction limit with respect of terrestrial controls. Eleven protein structures could be determined at previously unachieved resolutions using crystals obtained in the APCF. Microgravity induced features of the most relevant structures are reported. A second goal of this study was to identify the cause of the crystal quality enhancement useful for structure determination. No correlations between the effect of microgravity and other system-dependent parameters, such as isoelectric point or crystal solvent content, were found except the reduced convection during the crystallisation process. Thus, crystal growth under diffusive regime appears to be the key parameter explaining the beneficial effect of microgravity on crystal quality. The mimicry of these effects on earth in gels or in capillary tubes is discussed and the practical consequences for structural biology highlighted.},
note = {0301-4622 (Print)
Journal Article
Review},
keywords = {FRUGIER, GIEGE Animals *Biological Sciences Crystallization Equipment Design Humans Protein Conformation Proteins/*chemistry Research Support, Non-U.S. Gov't X-Ray Diffraction, SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Moreno A, Théobald-Dietrich A, Lorber B, Sauter C, Giege R
Effects of macromolecular impurities and of crystallization method on the quality of eubacterial aspartyl-tRNA synthetase crystals Article de journal
Dans: Acta Crystallogr D Biol Crystallogr, vol. 61, no. Pt 6, p. 789-792, 2005, ISBN: 15930641, (0907-4449 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: FRUGIER, SAUTER, Unité ARN
@article{,
title = {Effects of macromolecular impurities and of crystallization method on the quality of eubacterial aspartyl-tRNA synthetase crystals},
author = {A Moreno and A Théobald-Dietrich and B Lorber and C Sauter and R Giege},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15930641},
isbn = {15930641},
year = {2005},
date = {2005-01-01},
journal = {Acta Crystallogr D Biol Crystallogr},
volume = {61},
number = {Pt 6},
pages = {789-792},
abstract = {Although macromolecular purity is thought to be essential for the growth of flawless protein crystals, only a few studies have investigated how contaminants alter the crystallization process and crystal quality. Likewise, the outcome of a crystallization process may vary with the crystallization method. Here, it is reported how these two variables affect the crystallogenesis of aspartyl-tRNA synthetase from the eubacterium Thermus thermophilus. This homodimeric enzyme (Mr=130,000) possesses a multi-domain architecture and crystallizes either in a monoclinic or an orthorhombic habit. Minute amounts of protein impurities alter to a different extent the growth of each crystal form. The best synthetase crystals are only obtained when the crystallizing solution is either enclosed in capillaries or immobilized in agarose gel. In these two environments convection is reduced with regard to that existing in an unconstrained solution.},
note = {0907-4449
Journal Article},
keywords = {FRUGIER, SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Biertümpfel C, Basquin J, Birkenbihl R, Suck D, Sauter C
Characterization of crystals of the Hjc resolvase from Archaeoglobus fulgidus grown in gel by counter-diffusion. Article de journal
Dans: Acta Crystallogr F Struct Biol Commun, vol. 61, no. 7, p. 684-687, 2005.
Résumé | Liens | BibTeX | Étiquettes: FRUGIER, GIEGE Holliday junction-cutting enzyme Hjc Resolvase Counter-duffusion Agarose gel, SAUTER, Unité ARN
@article{,
title = {Characterization of crystals of the Hjc resolvase from Archaeoglobus fulgidus grown in gel by counter-diffusion.},
author = {C Biertümpfel and J Basquin and R Birkenbihl and D Suck and C Sauter},
url = {http://scripts.iucr.org/cgi-bin/paper?za5109},
doi = {10.1107/S1744309105018269},
year = {2005},
date = {2005-01-01},
journal = {Acta Crystallogr F Struct Biol Commun},
volume = {61},
number = {7},
pages = {684-687},
abstract = {Holliday junction-resolving enzymes are ubiquitous proteins that play a key role in DNA repair and reorganization by homologous recombination. The Holliday junction-cutting enzyme (Hjc) from the archaeon Archaeoglobus fulgidus is a member of this group. The first Hjc crystals were obtained by conventional sparse-matrix screening. They exhibited an unusually elongated unit cell and their X-ray characterization required special care to avoid spot overlaps along the c* axis. The use of an arc appended to the goniometric head allowed proper orientatation of plate-like crystals grown in agarose gel by counter-diffusion. Thus, complete diffraction data were collected at 2.7 Å resolution using synchrotron radiation. They belong to space group P3121 or P3221, with unit-cell parameters a = b = 37.4},
keywords = {FRUGIER, GIEGE Holliday junction-cutting enzyme Hjc Resolvase Counter-duffusion Agarose gel, SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
2004
Deniziak M A, Sauter C, Becker H D, Giege R, Kern D
Crystallization and preliminary X-ray characterization of the atypical glutaminyl-tRNA synthetase from Deinococcus radiodurans Article de journal
Dans: Acta Crystallogr D Biol Crystallogr, vol. 60, no. Pt 12 Pt 2, p. 2361-2363, 2004, ISBN: 15572799, (0907-4449 Journal article).
Résumé | Liens | BibTeX | Étiquettes: FRUGIER, KERN GIEGE, SAUTER, Unité ARN
@article{,
title = {Crystallization and preliminary X-ray characterization of the atypical glutaminyl-tRNA synthetase from Deinococcus radiodurans},
author = {M A Deniziak and C Sauter and H D Becker and R Giege and D Kern},
url = {http://journals.iucr.org/d/issues/2004/12/02/za0130/za0130bdy.html},
isbn = {15572799},
year = {2004},
date = {2004-01-01},
journal = {Acta Crystallogr D Biol Crystallogr},
volume = {60},
number = {Pt 12 Pt 2},
pages = {2361-2363},
abstract = {The glutaminyl-tRNA synthetase (GlnRS) from the radiation-resistant bacterium Deinococcus radiodurans differs from known GlnRSs and other tRNA synthetases by the presence of an additional C-terminal domain resembling the C-terminal region of the GatB subunit of tRNA-dependent amidotransferase (AdT). This atypical synthetase was overexpressed in Escherichia coli, purified and crystallized in the presence of PEG 3350. Orthorhombic crystals were obtained that belong to space group P2(1)2(1)2(1) and diffract to 2.3 A resolution. The crystal structure was solved by molecular replacement using the structure of E. coli GlnRS as a search model.},
note = {0907-4449
Journal article},
keywords = {FRUGIER, KERN GIEGE, SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
2003
Thore S, Mayer C, Sauter C, Weeks S, Suck D
Crystal structures of the Pyrococcus abyssi Sm core and its complex with RNA. Common features of RNA binding in Archaea and Eucarya. Article de journal
Dans: J Biol Chem, vol. 278, no. 2, p. 1239-1247, 2003, ISBN: 12409299.
Résumé | Liens | BibTeX | Étiquettes: FRUGIER, SAUTER, Unité ARN
@article{,
title = {Crystal structures of the Pyrococcus abyssi Sm core and its complex with RNA. Common features of RNA binding in Archaea and Eucarya.},
author = {S Thore and C Mayer and C Sauter and S Weeks and D Suck},
url = {http://www.ncbi.nlm.nih.gov/pubmed/12409299},
isbn = {12409299},
year = {2003},
date = {2003-01-01},
journal = {J Biol Chem},
volume = {278},
number = {2},
pages = {1239-1247},
abstract = {The Sm proteins are conserved in all three domains of life and are always associated with U-rich RNA sequences. Their proposed function is to mediate RNA-RNA interactions. We present here the crystal structures of Pyrococcus abyssi Sm protein (PA-Sm1) and its complex with a uridine heptamer. The overall structure of the protein complex, a heptameric ring with a central cavity, is similar to that proposed for the eukaryotic Sm core complex and found for other archaeal Sm proteins. RNA molecules bind to the protein at two different sites. They interact specifically inside the ring with three highly conserved residues, defining the uridine-binding pocket. In addition, nucleotides also interact on the surface formed by the N-terminal alpha-helix as well as a conserved aromatic residue in beta-strand 2 of the PA-Sm1 protein. The mutation of this conserved aromatic residue shows the importance of this second site for the discrimination between RNA sequences. Given the high structural homology between archaeal and eukaryotic Sm proteins, the PA-Sm1.RNA complex provides a model for how the small nuclear RNA contacts the Sm proteins in the Sm core. In addition, it suggests how Sm proteins might exert their function as modulators of RNA-RNA interactions.},
keywords = {FRUGIER, SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Sauter C, Basquin J, Suck D
Sm-like proteins in Eubacteria: the crystal structure of the Hfq protein from Escherichia coli Article de journal
Dans: Nucleic Acids Res, vol. 31, no. 14, p. 4091-4098, 2003, ISBN: 12853626, (1362-4962 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Amino Acid Support, Amino Acid Sequence Bacteria/*genetics Crystallography, FRUGIER, Molecular Host Factor 1 Protein/chemistry/*genetics Molecular Sequence Data Protein Conformation Protein Structure, Non-U.S. Gov't, SAUTER, Secondary Ribonucleoproteins, Small Nuclear/chemistry/*genetics Sequence Alignment Sequence Homology, Unité ARN, X-Ray Dimerization Escherichia coli Proteins/chemistry/*genetics Evolution
@article{,
title = {Sm-like proteins in Eubacteria: the crystal structure of the Hfq protein from Escherichia coli},
author = {C Sauter and J Basquin and D Suck},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12853626},
isbn = {12853626},
year = {2003},
date = {2003-01-01},
journal = {Nucleic Acids Res},
volume = {31},
number = {14},
pages = {4091-4098},
abstract = {The Hfq protein was discovered in Escherichia coli in the early seventies as a host factor for the Qbeta phage RNA replication. During the last decade, it was shown to be involved in many RNA processing events and remote sequence homology indicated a link to spliceosomal Sm proteins. We report the crystal structure of the E.coli Hfq protein showing that its monomer displays a characteristic Sm-fold and forms a homo-hexamer, in agreement with former biochemical data. Overall, the structure of the E.coli Hfq ring is similar to the one recently described for Staphylococcus aureus. This confirms that bacteria contain a hexameric Sm-like protein which is likely to be an ancient and less specialized form characterized by a relaxed RNA binding specificity. In addition, we identified an Hfq ortholog in the archaeon Methanococcus jannaschii which lacks a classical Sm/Lsm gene. Finally, a detailed structural comparison shows that the Sm-fold is remarkably well conserved in bacteria, Archaea and Eukarya, and represents a universal and modular building unit for oligomeric RNA binding proteins.},
note = {1362-4962
Journal Article},
keywords = {Amino Acid Support, Amino Acid Sequence Bacteria/*genetics Crystallography, FRUGIER, Molecular Host Factor 1 Protein/chemistry/*genetics Molecular Sequence Data Protein Conformation Protein Structure, Non-U.S. Gov't, SAUTER, Secondary Ribonucleoproteins, Small Nuclear/chemistry/*genetics Sequence Alignment Sequence Homology, Unité ARN, X-Ray Dimerization Escherichia coli Proteins/chemistry/*genetics Evolution},
pubstate = {published},
tppubtype = {article}
}
2002
Biertümpfel C, Basquin J, Suck D, Sauter C
Crystallization of biological macromolecules using agarose gel. Article de journal
Dans: Acta Crystallogr D Biol Crystallogr, vol. 58, no. 10, p. 1657-1659, 2002, ISBN: 10.1107/S0907444902012738.
Résumé | Liens | BibTeX | Étiquettes: Agarose gel Crystallization Biological macromolecule Counter-diffusion., FRUGIER, SAUTER, Unité ARN
@article{,
title = {Crystallization of biological macromolecules using agarose gel.},
author = {C Biertümpfel and J Basquin and D Suck and C Sauter},
url = {http://scripts.iucr.org/cgi-bin/paper?ic0008},
isbn = {10.1107/S0907444902012738},
year = {2002},
date = {2002-01-01},
journal = {Acta Crystallogr D Biol Crystallogr},
volume = {58},
number = {10},
pages = {1657-1659},
abstract = {Gellified media prevent convection and crystal sedimentation, and provide an attractive growth environment for optimising biological crystals. Agarose gels are particularly easy to use and they are compatible with most of the common crystallization methods. They also offer new possibilities like counter-diffusion techniques. This paper gives a brief overview of their general properties and presents an application of a counter-diffusion setup combining agarose gel and capillaries to the crystallization of proteins and protein / nucleic acid complexes.},
keywords = {Agarose gel Crystallization Biological macromolecule Counter-diffusion., FRUGIER, SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Sauter C, Lorber B, Giege R
Towards atomic resolution with crystals grown in gel: the case of thaumatin seen at room temperature Article de journal
Dans: Proteins, vol. 48, no. 2, p. 146-150, 2002, ISBN: 12112683, (1097-0134 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Crystallization *Crystallography, FRUGIER, Molecular Plant Proteins/*chemistry Support, Non-U.S. Gov't *Sweetening Agents Temperature Weightlessness, SAUTER, Unité ARN, X-Ray Hydrogels/*chemistry *Models
@article{,
title = {Towards atomic resolution with crystals grown in gel: the case of thaumatin seen at room temperature},
author = {C Sauter and B Lorber and R Giege},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12112683},
isbn = {12112683},
year = {2002},
date = {2002-01-01},
journal = {Proteins},
volume = {48},
number = {2},
pages = {146-150},
abstract = {One reason for introducing a gel in the crystallization medium of proteins is its ability to reduce convection in solution. This can lead to better nucleation and growth conditions, and to crystals having enhanced diffraction properties. We report here the X-ray characterization at room temperature of high-quality crystals of the intensely sweet thaumatin prepared in a sodium tartrate solution gelified with 0.15% (m/v) agarose. Using a synchrotron radiation, these crystals diffracted to a previously unachieved resolution. A diffraction dataset was collected from four crystals at a resolution of 1.2 A with a R(sym) of 3.6% and a completeness of 99%. Refinement was carried out to a final crystallographic R-factor of 12.0%. The quality of the electron density map allowed for the observation of fine structural details in the protein and its solvation shell. Crystallization in gel might be used more generally to improve the quality of macromolecular crystals. Advantages provided by the gelified medium in the frame of structural studies are emphasized.},
note = {1097-0134
Journal Article},
keywords = {Crystallization *Crystallography, FRUGIER, Molecular Plant Proteins/*chemistry Support, Non-U.S. Gov't *Sweetening Agents Temperature Weightlessness, SAUTER, Unité ARN, X-Ray Hydrogels/*chemistry *Models},
pubstate = {published},
tppubtype = {article}
}
Ng J D, Sauter C, Lorber B, Kirkland N, Arnez J, Giege R
Comparative analysis of space-grown and earth-grown crystals of an aminoacyl-tRNA synthetase: space-grown crystals are more useful for structural determination Article de journal
Dans: Acta Crystallogr D Biol Crystallogr, vol. 58, no. Pt 4, p. 645-652, 2002, ISBN: 11914489, (0907-4449 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Amino Acyl-tRNA Ligases/*chemistry Comparative Study Crystallization Crystallography, FRUGIER, Molecular Support, Non-U.S. Gov't Thermus thermophilus/chemistry Water/chemistry Weightlessness, SAUTER, Unité ARN, X-Ray Models
@article{,
title = {Comparative analysis of space-grown and earth-grown crystals of an aminoacyl-tRNA synthetase: space-grown crystals are more useful for structural determination},
author = {J D Ng and C Sauter and B Lorber and N Kirkland and J Arnez and R Giege},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11914489},
isbn = {11914489},
year = {2002},
date = {2002-01-01},
journal = {Acta Crystallogr D Biol Crystallogr},
volume = {58},
number = {Pt 4},
pages = {645-652},
abstract = {Protein crystallization under microgravity aims at benefiting from the quasi-absence of convection and sedimentation to favor well ordered crystal nucleation and growth. The dimeric multidomain enzyme aspartyl-tRNA synthetase from Thermus thermophilus has been crystallized within dialysis reactors of the Advanced Protein Crystallization Facility in the laboratory on earth and under microgravity aboard the US Space Shuttle. A strictly comparative crystallographic analysis reveals that the crystals grown in space are superior in every respect to control crystals prepared in otherwise identical conditions on earth. They diffract X-rays more intensely and have a lower mosaicity, facilitating the process of protein structure determination. Indeed, the electron-density map calculated from diffraction data of space-grown crystals contains considerably more detail. The resulting three-dimensional structure model at 2.0 A resolution is more accurate than that produced in parallel using the data originating from earth-grown crystals. The major differences between the structures, including the better defined amino-acid side chains and the higher order of bound water molecules, are emphasized.},
note = {0907-4449
Journal Article},
keywords = {Amino Acyl-tRNA Ligases/*chemistry Comparative Study Crystallization Crystallography, FRUGIER, Molecular Support, Non-U.S. Gov't Thermus thermophilus/chemistry Water/chemistry Weightlessness, SAUTER, Unité ARN, X-Ray Models},
pubstate = {published},
tppubtype = {article}
}
Lorber B, Théobald-Dietrich A, Charron C, Sauter C, Ng J D, Zhu D W, Giege R
From conventional crystallization to better crystals from space: a review on pilot crystallogenesis studies with aspartyl-tRNA synthetases Article de journal
Dans: Acta Crystallogr D Biol Crystallogr, vol. 58, no. Pt 10 Pt 1, p. 1674-1680, 2002, ISBN: 12351885, (0907-4449 Journal Article Review Review, Tutorial).
Résumé | Liens | BibTeX | Étiquettes: Aspartate-tRNA Ligase/*chemistry Crystallization/*methods Crystallography, Non-U.S. Gov't Thermus thermophilus/enzymology Weightlessness, SAUTER, Unité ARN, X-Ray Molecular Structure Pilot Projects Saccharomyces cerevisiae/enzymology Space Flight Support
@article{,
title = {From conventional crystallization to better crystals from space: a review on pilot crystallogenesis studies with aspartyl-tRNA synthetases},
author = {B Lorber and A Théobald-Dietrich and C Charron and C Sauter and J D Ng and D W Zhu and R Giege},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12351885},
isbn = {12351885},
year = {2002},
date = {2002-01-01},
journal = {Acta Crystallogr D Biol Crystallogr},
volume = {58},
number = {Pt 10 Pt 1},
pages = {1674-1680},
abstract = {Aspartyl-tRNA synthetases were the model proteins in pilot crystallogenesis experiments. They are homodimeric enzymes of Mr approximately 125 kDa that possess as substrates a transfer RNA, ATP and aspartate. They have been isolated from different sources and were crystallized either as free proteins or in association with their ligands. This review discusses their crystallisability with emphasis to crystal quality and structure determination. Crystallization in low diffusivity gelled media or in microgravity environments is highlighted. It has contributed to prepare high-resolution diffracting crystals with better internal order as reflected by their mosaicity. With AspRS from Thermus thermophilus, the better crystalline quality of the space-grown crystals within APCF is correlated with higher quality of the derived electron density maps. Usefulness for structural biology of targeted methods aimed to improve the intrinsic physical quality of protein crystals is highlighted.},
note = {0907-4449
Journal Article
Review
Review, Tutorial},
keywords = {Aspartate-tRNA Ligase/*chemistry Crystallization/*methods Crystallography, Non-U.S. Gov't Thermus thermophilus/enzymology Weightlessness, SAUTER, Unité ARN, X-Ray Molecular Structure Pilot Projects Saccharomyces cerevisiae/enzymology Space Flight Support},
pubstate = {published},
tppubtype = {article}
}
2001
Sauter C, Otalora F, Gavira J A, Vidal O, Giege R, Garcia-Ruiz J M
Structure of tetragonal hen egg-white lysozyme at 0.94 A from crystals grown by the counter-diffusion method Article de journal
Dans: Acta Crystallogr D Biol Crystallogr, vol. 57, no. Pt 8, p. 1119-1126, 2001, ISBN: 11468395, (0907-4449 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Amino Acids/chemistry Animals Anions/chemistry Binding Sites Cations/chemistry Chickens Crystallization Crystallography, Molecular Muramidase/*chemistry Protein Conformation Protein Structure, Non-U.S. Gov't, SAUTER, Tertiary Support, Unité ARN, X-Ray Egg White/analysis Models
@article{,
title = {Structure of tetragonal hen egg-white lysozyme at 0.94 A from crystals grown by the counter-diffusion method},
author = {C Sauter and F Otalora and J A Gavira and O Vidal and R Giege and J M Garcia-Ruiz},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11468395},
isbn = {11468395},
year = {2001},
date = {2001-01-01},
journal = {Acta Crystallogr D Biol Crystallogr},
volume = {57},
number = {Pt 8},
pages = {1119-1126},
abstract = {Very high quality crystals of tetragonal hen egg-white lysozyme were grown in the Advanced Protein Crystallization Facility (APCF) on board the Space Shuttle using a modified free-interface diffusion (FID) reactor designed ad hoc to have a longer diffusion path. This design allows the performance of true counter-diffusion experiments. Crystals were obtained under the classical chemical conditions defined 50 y ago with NaCl as a crystallizing agent and acetate pH 4.5 as a buffer. Counter-diffusion crystallization allows a "physical" instead of chemical optimization of growth conditions: indeed, this method screens for the best supersaturation conditions in a single trial and yields crystals of very high quality. A complete diffraction data set was collected at atomic resolution from one of these crystals using synchrotron radiation at the DESY-EMBL beamlines. The overall R(merge) on intensities in the resolution range 31-0.94 A was 5.2% and the data were 98.9% complete. Refinement was carried out with the programs CNS and SHELX97 to a final crystallographic R factor of 12.26% for 72 390 reflections. A mean standard uncertainty in the atomic positions of 0.024 A was estimated from inversion of blocked least-squares matrices. 22 side chains show alternate conformations and the loop 59-75 adopts in the same crystal packing two conformations that were observed for either triclinic or tetragonal lysozyme in previous high-resolution studies. In addition to 255 water molecules, the crystallizing agent (one hexacoordinated sodium ion and five chloride anions) participates in the ordered lysozyme hydration shell.},
note = {0907-4449
Journal Article},
keywords = {Amino Acids/chemistry Animals Anions/chemistry Binding Sites Cations/chemistry Chickens Crystallization Crystallography, Molecular Muramidase/*chemistry Protein Conformation Protein Structure, Non-U.S. Gov't, SAUTER, Tertiary Support, Unité ARN, X-Ray Egg White/analysis Models},
pubstate = {published},
tppubtype = {article}
}
Zhu D W, Lorber B, Sauter C, Ng J D, Benas P, Grimellec C Le, Giege R
Growth kinetics, diffraction properties and effect of agarose on the stability of a novel crystal form of Thermus thermophilus aspartyl-tRNA synthetase-1 Article de journal
Dans: Acta Crystallogr D Biol Crystallogr, vol. 57, no. Pt 4, p. 552-558, 2001, ISBN: 11264584, (0907-4449 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Aspartate-tRNA Ligase/*chemistry/*metabolism Crystallization Crystallography, Atomic Force Osmolar Concentration Sepharose/*metabolism Solubility Support, Non-U.S. Gov't Temperature Thermodynamics Thermus thermophilus/*enzymology, SAUTER, Unité ARN, X-Ray/methods Enzyme Stability Gels Kinetics Microscopy
@article{,
title = {Growth kinetics, diffraction properties and effect of agarose on the stability of a novel crystal form of Thermus thermophilus aspartyl-tRNA synthetase-1},
author = {D W Zhu and B Lorber and C Sauter and J D Ng and P Benas and C Le Grimellec and R Giege},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11264584},
isbn = {11264584},
year = {2001},
date = {2001-01-01},
journal = {Acta Crystallogr D Biol Crystallogr},
volume = {57},
number = {Pt 4},
pages = {552-558},
abstract = {Growth kinetics and diffraction properties of monoclinic crystals of eubacterial Thermus thermophilus aspartyl-tRNA synthetase-1 (AspRS-1) prepared in the presence of polyethylene glycol and agarose are studied. Their solubility and two-dimensional phase diagram are compared with those of orthorhombic crystals which grow in the presence of sodium formate or ammonium sulfate. The growth mechanism of the novel crystals was monitored by atomic force microscopy. The gel stabilizes the crystal lattice under the cryogenic conditions used for structure determination at high resolution.},
note = {0907-4449
Journal Article},
keywords = {Aspartate-tRNA Ligase/*chemistry/*metabolism Crystallization Crystallography, Atomic Force Osmolar Concentration Sepharose/*metabolism Solubility Support, Non-U.S. Gov't Temperature Thermodynamics Thermus thermophilus/*enzymology, SAUTER, Unité ARN, X-Ray/methods Enzyme Stability Gels Kinetics Microscopy},
pubstate = {published},
tppubtype = {article}
}
Théobald-Dietrich A, Lorber B, Ng J D, Sauter C, Charron C, Robert M C, Capelle B, Giege R
Crystals grown in microgravity can yield better protein structures: the examples of thaumatin and aspartyl-tRNA synthetase. Chapitre d'ouvrage
Dans: Minster, O; Schurmann, B (Ed.): Microgravity Research and Aplications in Physical Sciences and Biotechnology, Proceedings of the First International Symposium held 10-15 September, 2000 in Sorrento, Italy., p. 457-463, European Space Agency, 2001.
Liens | BibTeX | Étiquettes: SAUTER, Unité ARN
@inbook{,
title = {Crystals grown in microgravity can yield better protein structures: the examples of thaumatin and aspartyl-tRNA synthetase.},
author = {A Théobald-Dietrich and B Lorber and J D Ng and C Sauter and C Charron and M C Robert and B Capelle and R Giege},
editor = {O Minster and B Schurmann},
url = {http://adsabs.harvard.edu/abs/2001ESASP.454.457T},
year = {2001},
date = {2001-01-01},
booktitle = {Microgravity Research and Aplications in Physical Sciences and Biotechnology, Proceedings of the First International Symposium held 10-15 September, 2000 in Sorrento, Italy.},
pages = {457-463},
publisher = {European Space Agency},
keywords = {SAUTER, Unité ARN},
pubstate = {published},
tppubtype = {inbook}
}