Intranet
Acces
Contact
Publications
2002
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}
}
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.
2000
Sauter C, Lorber B, Cavarelli J, Moras D, Giege R
The free yeast aspartyl-tRNA synthetase differs from the tRNA(Asp)-complexed enzyme by structural changes in the catalytic site, hinge region, and anticodon-binding domain Article de journal
Dans: J Mol Biol, vol. 299, no. 5, p. 1313-1324, 2000, ISBN: 10873455, (0022-2836 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Anticodon/chemistry/genetics/*metabolism Aspartate-tRNA Ligase/*chemistry/genetics/*metabolism Binding Sites Catalytic Domain Conserved Sequence/genetics Crystallization Crystallography, Asp/chemistry/genetics/*metabolism Rotation Sequence Deletion/genetics Support, Fungal/chemistry/genetics/metabolism RNA, Molecular Molecular Sequence Data Movement Nucleic Acid Conformation Protein Structure, Non-U.S. Gov't Yeasts/*enzymology/genetics, SAUTER, Secondary RNA, Transfer, Unité ARN, X-Ray Models
@article{,
title = {The free yeast aspartyl-tRNA synthetase differs from the tRNA(Asp)-complexed enzyme by structural changes in the catalytic site, hinge region, and anticodon-binding domain},
author = {C Sauter and B Lorber and J Cavarelli and D Moras and R Giege},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10873455},
isbn = {10873455},
year = {2000},
date = {2000-01-01},
journal = {J Mol Biol},
volume = {299},
number = {5},
pages = {1313-1324},
abstract = {Aminoacyl-tRNA synthetases catalyze the specific charging of amino acid residues on tRNAs. Accurate recognition of a tRNA by its synthetase is achieved through sequence and structural signalling. It has been shown that tRNAs undergo large conformational changes upon binding to enzymes, but little is known about the conformational rearrangements in tRNA-bound synthetases. To address this issue the crystal structure of the dimeric class II aspartyl-tRNA synthetase (AspRS) from yeast was solved in its free form and compared to that of the protein associated to the cognate tRNA(Asp). The use of an enzyme truncated in N terminus improved the crystal quality and allowed us to solve and refine the structure of free AspRS at 2.3 A resolution. For the first time, snapshots are available for the different macromolecular states belonging to the same tRNA aminoacylation system, comprising the free forms for tRNA and enzyme, and their complex. Overall, the synthetase is less affected by the association than the tRNA, although significant local changes occur. They concern a rotation of the anticodon binding domain and a movement in the hinge region which connects the anticodon binding and active-site domains in the AspRS subunit. The most dramatic differences are observed in two evolutionary conserved loops. Both are in the neighborhood of the catalytic site and are of importance for ligand binding. The combination of this structural analysis with mutagenesis and enzymology data points to a tRNA binding process that starts by a recognition event between the tRNA anticodon loop and the synthetase anticodon binding module.},
note = {0022-2836
Journal Article},
keywords = {Anticodon/chemistry/genetics/*metabolism Aspartate-tRNA Ligase/*chemistry/genetics/*metabolism Binding Sites Catalytic Domain Conserved Sequence/genetics Crystallization Crystallography, Asp/chemistry/genetics/*metabolism Rotation Sequence Deletion/genetics Support, Fungal/chemistry/genetics/metabolism RNA, Molecular Molecular Sequence Data Movement Nucleic Acid Conformation Protein Structure, Non-U.S. Gov't Yeasts/*enzymology/genetics, SAUTER, Secondary RNA, Transfer, Unité ARN, X-Ray Models},
pubstate = {published},
tppubtype = {article}
}
Aminoacyl-tRNA synthetases catalyze the specific charging of amino acid residues on tRNAs. Accurate recognition of a tRNA by its synthetase is achieved through sequence and structural signalling. It has been shown that tRNAs undergo large conformational changes upon binding to enzymes, but little is known about the conformational rearrangements in tRNA-bound synthetases. To address this issue the crystal structure of the dimeric class II aspartyl-tRNA synthetase (AspRS) from yeast was solved in its free form and compared to that of the protein associated to the cognate tRNA(Asp). The use of an enzyme truncated in N terminus improved the crystal quality and allowed us to solve and refine the structure of free AspRS at 2.3 A resolution. For the first time, snapshots are available for the different macromolecular states belonging to the same tRNA aminoacylation system, comprising the free forms for tRNA and enzyme, and their complex. Overall, the synthetase is less affected by the association than the tRNA, although significant local changes occur. They concern a rotation of the anticodon binding domain and a movement in the hinge region which connects the anticodon binding and active-site domains in the AspRS subunit. The most dramatic differences are observed in two evolutionary conserved loops. Both are in the neighborhood of the catalytic site and are of importance for ligand binding. The combination of this structural analysis with mutagenesis and enzymology data points to a tRNA binding process that starts by a recognition event between the tRNA anticodon loop and the synthetase anticodon binding module.
