RNA interference and receptors

@article{bianco_solid-phase_2006,

title = {Solid-phase synthesis of CD40L mimetics},

author = {Alberto Bianco and Sylvie Fournel and Sébastien Wieckowski and Johan Hoebeke and Gilles Guichard},

doi = {10.1039/b601528j},

issn = {1477-0520},

year  = {2006},

date = {2006-04-01},

journal = {Organic & Biomolecular Chemistry},

volume = {4},

number = {8},

pages = {1461--1463},

abstract = {The C3-symmetric molecule has been previously shown to mimic CD40 ligand (CD40L) homotrimers and to display effector functions. This molecule consists of a cyclic hexapeptide core containing the repetition of the D-Ala-L-Lys motif. The side chains of the lysine residues have been modified by appending the CD40L-derived sequence 143Lys-Gly-Tyr-Tyr146 via a 6-aminohexanoic acid residue as a spacer. The present report describes a general solid-phase synthesis approach to and related trimeric architectures. In addition, their CD40 binding properties as well as their effector functions have been evaluated.},

keywords = {Apoptosis, CD40 Ligand, Cell Line, Chromatography, Combinatorial Chemistry Techniques, High Pressure Liquid, Humans, I2CT, Molecular Mimicry, Molecular Structure, Protein Binding, Team-Bianco, tumor},

pubstate = {published},

tppubtype = {article}

}

@article{brough_60fullerene-pyrrolidine-n-oxides_2006,

title = {[60]fullerene-pyrrolidine-N-oxides},

author = {Peter Brough and Cedric Klumpp and Alberto Bianco and Stephane Campidelli and Maurizio Prato},

doi = {10.1021/jo052388s},

issn = {0022-3263},

year  = {2006},

date = {2006-03-01},

journal = {The Journal of Organic Chemistry},

volume = {71},

number = {5},

pages = {2014--2020},

abstract = {Eight members of a new family of fullerene derivatives, [60]fulleropyrrolidine-N-oxides, have been synthesized and characterized. Facile oxidation, by a peracid, of the parent [60]fulleropyrrolidine gave clean conversions into the product molecules, in which the tertiary amine is transformed into a quaternary amine bearing an oxygen atom. The reaction is very selective, favoring the nitrogen atom of the pyrrolidine ring in preference to epoxidation of the fullerene cage. The 1H NMR shows an AB quartet splitting pattern, characteristic of nonequivalent hydrogens in the pyrrolidine ring and at a chemical shift displacement of 0.8 ppm downfield. Other methods of characterization are described, including MS, differential scanning calorimetry, thermogravimetric analysis, HPLC, UV/vis, and IR. Conclusive evidence for the formation of an N-oxide moiety is provided by the synthesis, oxidation, and NMR characterization of a novel [60]fulleropyrrolidine containing a 15N isotope, showing an 85 ppm downfield heteroatom chemical shift. Preliminary details of the effects of substitution on the reactivity of the pyrrolidine ring are also reported.},

keywords = {Chromatography, Fullerenes, High Pressure Liquid, I2CT, Nitrogen, Oxidation-Reduction, Oxides, Pyrrolidines, Spectrum Analysis, Team-Bianco},

pubstate = {published},

tppubtype = {article}

}

@article{bianco_efficient_2005,

title = {Efficient solid-phase synthesis of fullero-peptides using Merrifield strategy},

author = {Alberto Bianco},

doi = {10.1039/b504659a},

issn = {1359-7345},

year  = {2005},

date = {2005-07-01},

journal = {Chemical Communications (Cambridge, England)},

number = {25},

pages = {3174--3176},

abstract = {Boc-protected l-fulleropyrrolidino-glutamic acid was readily prepared and employed for the synthesis of fullerene-containing peptides using the solid-phase Boc chemistry developed by Merrifield.},

keywords = {Chromatography, Fullerenes, High Pressure Liquid, I2CT, Mass Spectrometry, Peptides, Team-Bianco},

pubstate = {published},

tppubtype = {article}

}

@article{,

title = {A simple epigenetic method for the diagnosis and classification of brain tumors},

author = { R. Zukiel and S. Nowak and A. M. Barciszewska and I. Gawronska and G. Keith and M. Z. Barciszewska},

year  = {2004},

date = {2004-01-01},

journal = {Mol Cancer Res},

volume = {2},

number = {3},

pages = {196-202},

abstract = {The new, simple, and reliable method for the diagnosis of brain tumors is described. It is based on a TLC quantitative determination of 5-methylcytosine (m(5)C) in relation to its damage products of DNA from tumor tissue. Currently, there is evidence that oxidative stress through reactive oxygen species (ROS) plays an important role in the etiology and progression of several human diseases. Oxidative damage of DNA, lipids, and proteins is deleterious for the cell. m(5)C, along with other basic components of DNA, is the target for ROS, which results in the appearance of new modified nucleic acid bases. If so, m(5)C residue constitutes a mutational hotspot position, whether it occurs within a nucleotide sequence of a structural gene or a regulatory region. Here, we show the results of the analysis of 82 DNA samples taken from brain tumor tissues. DNA was isolated and hydrolyzed into nucleotides, which, after labeling with [gamma-(32)P]ATP, were separated on TLC. Chromatograms were evaluated using PhosphorImager and the amounts of 5-methyldeoxycytosine (m(5)dC) were calculated as a ratio (R) of m(5)dC to m(5)dC + deoxycytosine + deoxythymidine spot intensities. The R value could not only be a good diagnostic marker for brain tumors but also a factor differentiating low-grade and high-grade gliomas. Therefore, DNA methylation pattern might be a useful tool to give a primary diagnosis of a brain tumor or as a marker for the early detection of the relapse of the disease. This method has several advantages over those existing nowadays.},

note = {1541-7786 

Journal Article},

keywords = {*DNA, *Epigenesis, 5-Methylcytosine/*analysis, Adult, Aged, and, Brain, Chromatography, DNA, Female, Genetic, Gov't, Human, KEITH, Layer, Male, Methylation, Middle, Neoplasm/*chemistry/*metabolism, Neoplasms/*classification/*diagnosis/genetics/pathology, Non-U.S., Oxidative, oxygen, Reactive, Sensitivity, Species/metabolism, Specificity, Stress, Support, Thin},

pubstate = {published},

tppubtype = {article}

}

@article{bianco_solid-phase_2003,

title = {Solid-phase synthesis and characterization of a novel fullerene-peptide derived from histone H3},

author = {Alberto Bianco and Davide Pantarotto and Johan Hoebeke and Jean-Paul Briand and Maurizio Prato},

doi = {10.1039/b311505d},

issn = {1477-0520},

year  = {2003},

date = {2003-12-01},

journal = {Organic & Biomolecular Chemistry},

volume = {1},

number = {23},

pages = {4141--4143},

abstract = {A peptide analogue from a histone H3 protein containing the L-fulleropyrrolidino-glutamic acid has been prepared by a solid-phase approach and has been fully characterized. By molecular modelling it was verified that this peptide derivative is able to retain a binding capacity to the MHC (major histocompatibility complex) molecule similar to that of the cognate epitope.},

keywords = {Chromatography, Epitopes, Fullerenes, Glutamic Acid, High Pressure Liquid, Histones, I2CT, Models, Molecular, Molecular Structure, Peptides, Protein Structure, Team-Bianco, Tertiary},

pubstate = {published},

tppubtype = {article}

}

@article{pantarotto_synthesis_2003,

title = {Synthesis, structural characterization, and immunological properties of carbon nanotubes functionalized with peptides},

author = {Davide Pantarotto and Charalambos D Partidos and Roland Graff and Johan Hoebeke and Jean-Paul Briand and Maurizio Prato and Alberto Bianco},

doi = {10.1021/ja034342r},

issn = {0002-7863},

year  = {2003},

date = {2003-05-01},

journal = {Journal of the American Chemical Society},

volume = {125},

number = {20},

pages = {6160--6164},

abstract = {Carbon nanotubes (NTs) are becoming highly attractive molecules for applications in medicinal chemistry. The main problem of insolubility in aqueous media has been solved by developing a synthetic protocol that allows highly water-soluble carbon NTs to be obtained. As a result, biologically active peptides can be easily linked through a stable covalent bond to carbon NTs. We have demonstrated that a bound peptide from the foot-and-mouth disease virus, corresponding to the 141-159 region of the viral envelope protein VP1, retained the structural integrity and was recognized by monoclonal and polyclonal antibodies. In addition, this peptide-NT conjugate is immunogenic, eliciting antibody responses of the right specificity. Such a system could be greatly advantageous for diagnostic purposes and could find future applications in vaccine delivery.},

keywords = {B-Lymphocyte, biomolecular, Capsid Proteins, carbon, Chromatography, Epitopes, Foot-and-Mouth Disease Virus, High Pressure Liquid, I2CT, nanotechnology, Nanotubes, Nuclear Magnetic Resonance, Peptide Fragments, Team-Bianco},

pubstate = {published},

tppubtype = {article}

}

@article{,

title = {Purification of turkey pancreatic phospholipase A2},

author = { R. Ben Salah and N. Zouari and J. Reinbolt and H. Mejdoub},

year  = {2003},

date = {2003-01-01},

journal = {Biosci Biotechnol Biochem},

volume = {67},

number = {10},

pages = {2139-44},

abstract = {Turkey pancreatic phospholipase (TPP) has been purified from delipidated pancreases. The purification included ammonium sulfate fractionation, acidic (pH 5) treatment, followed by sequencial column chromatographies on DEAE-cellulose, Sephadex G-75, and reverse phase high pressure liquid chromatography. The purified enzyme was found to be a monomeric protein with molecular mass of 14 kDa. The optimal activity was measured at pH 8 and 37 degrees C using egg yolk emulsion as substrate. Our results show that the enzyme (TPP) was not stable for 1 h at 60 degrees C, and that bile salt and Ca2+ were required for the expression of the purified enzyme. The sequence of the N-terminal amino acids of the purified enzyme shows a very close similarity between TPP and all other known pancreatic phospholipases.},

note = {0916-8451 

Journal Article},

keywords = {*Turkeys, &, A/*isolation, Acid, acids, Amino, Ammonium, and, Animals, Bile, Calcium, Chromatography, Concentration, Data, Hydrogen-Ion, Molecular, Pancreas/*enzymology, Phospholipases, purification, Salts, Sequence, Sulfate, Temperature, Weight},

pubstate = {published},

tppubtype = {article}

}

@article{,

title = {Lack of genotoxicity of bitumen fumes in transgenic mouse lung},

author = { J. C. Micillino and C. Coulais and S. Binet and M. C. Bottin and G. Keith and D. Moulin and B. H. Rihn},

year  = {2002},

date = {2002-01-01},

journal = {Toxicology},

volume = {170},

number = {1-2},

pages = {11-20},

abstract = {During hot application of bitumen containing materials, e.g. in hot paving or roofing, fumes are emitted that contain polycyclic aromatic compounds. Previous studies with rodents exposed to bitumen and coal-tar fume condensates showed formation of DNA adducts. In order to clarify the genotoxicity of bitumen fumes, we designed a study by using mice carrying a reporter gene for mutagenesis analysis and exposed by nose-only to a constant and reproducible aerosol of bitumen fumes. We analyzed the genotoxic activity of inhaled bitumen fumes generated under those controlled conditions through the induction of mutation and DNA adducts in Big Blue mice. Mice were exposed to bitumen fumes (100 mg/m(3) total particulate matter) 6 h per day during 5 days by nose-only in an inhalation chamber designed in our laboratory. Following a 30-day fixation period, the experiment was terminated and lung DNA was extracted for mutant frequency and adduct determinations. The mutant frequency was determined using the cII and the lacI mutant analysis systems. In, addition, 61 and 54 mutants were sequenced in control and exposed groups, respectively. The study did not show any mutation or adduct induction in the exposed group compared to the control group: cII mutant frequencies were 11.0+/-4.5x10(-5) and 11.0+/-4.8x10(-5) in control and exposed lungs, respectively. Identically, using the lacI mutation detection system, the mutant frequencies were 6.4+/-3.1x10(-5) and 5.8+/-2.0x10(-5). The mutation spectra of both series were quite similar with regard to transition and transversion frequencies. The absence of genotoxicity in the group exposed to 100 mg/m(3) bitumen is discussed with regard to dosage of inhaled polycyclic aromatic compounds and species.},

note = {0300-483x 

Journal Article},

keywords = {Adducts/drug, Aerosols, Animals, C57BL, Chromatography, DNA, DNA/drug, effects, effects/metabolism, Gases/*toxicity, Genes, Hydrocarbons/*toxicity, Inbred, Lac, Layer, Lung/*drug, Mice, Mutagenicity, Mutagens/*toxicity, Mutation/drug, Operon/genetics, Reporter/genetics, Tests, Thin, transgenic},

pubstate = {published},

tppubtype = {article}

}

@article{lamberty_insect_2001,

title = {Insect immunity. Constitutive expression of a cysteine-rich antifungal and a linear antibacterial peptide in a termite insect.},

author = {M Lamberty and Daniel Zachary and R Lanot and C Bordereau and A Robert and Jules A Hoffmann and Philippe Bulet},

doi = {10.1074/jbc.M002998200},

issn = {0021-9258},

year  = {2001},

date = {2001-02-01},

journal = {J. Biol. Chem.},

volume = {276},

number = {6},

pages = {4085--4092},

abstract = {Two novel antimicrobial peptides, which we propose to name termicin and spinigerin, have been isolated from the fungus-growing termite Pseudacanthotermes spiniger (heterometabole insect, Isoptera). Termicin is a 36-amino acid residue antifungal peptide, with six cysteines arranged in a disulfide array similar to that of insect defensins. In contrast to most insect defensins, termicin is C-terminally amidated. Spinigerin consists of 25 amino acids and is devoid of cysteines. It is active against bacteria and fungi. Termicin and spinigerin show no obvious sequence similarities with other peptides. Termicin is constitutively present in hemocyte granules and in salivary glands. The presence of termicin and spinigerin in unchallenged termites contrasts with observations in evolutionary recent insects or insects undergoing complete metamorphosis, in which antimicrobial peptides are induced in the fat body and released into the hemolymph after septic injury.},

keywords = {Amino Acid, Animals, Anti-Bacterial Agents, Antifungal Agents, Base Sequence, Chromatography, Cysteine, DNA Primers, High Pressure Liquid, hoffmann, Immunohistochemistry, Isoptera, M3i, Peptides, Protein Conformation, Recombinant Proteins, Sequence Homology},

pubstate = {published},

tppubtype = {article}

}

@article{adamkewicz_purification_2000,

title = {Purification and enzymic properties of Mot1 ATPase, a regulator of basal transcription in the yeast Saccharomyces cerevisiae},

author = {J I Adamkewicz and C G Mueller and K E Hansen and W A Prud'homme and J Thorner},

doi = {10.1074/jbc.M002639200},

issn = {0021-9258},

year  = {2000},

date = {2000-07-01},

journal = {The Journal of Biological Chemistry},

volume = {275},

number = {28},

pages = {21158--21168},

abstract = {The 1867-residue Mot1 protein is a member of a superfamily of ATPases, some of which are helicases, that interact with protein-nucleic acid assemblies. Mot1 is an essential regulator of RNA polymerase II-dependent transcription in vivo and dissociates TATA box-binding protein (TBP)-DNA complexes in vitro. Mot1-(His)(6) was purified to apparent homogeneity from yeast extracts. The preparation efficiently dissociated TBP.TATA complexes, suggesting that no other protein or cofactor is required. Mot1 behaved as a non-globular monomer in hydrodynamic studies, and no association was detected between differentially tagged co-expressed Mot1 constructs. ATPase activity was stimulated about 10-fold by high ionic strength or alkaline pH, or by deletion of the N-terminal TBP-binding segment, suggesting that the N-terminal domain negatively regulates the C-terminal ATPase domain (Mot1C). Correspondingly, at moderate salt concentration, Mot1 ATPase (but not Mot1C) was stimulated textgreater/=10-fold by yeast TBP, suggesting that interaction with TBP relieves a conformational constraint in Mot1. Double- or single-stranded TATA-containing DNA did not affect ATPase activity of Mot1 or Mot1C, with or without TBP. Mot1 did not exhibit detectable helicase activity in strand displacement assays using substrates with flush ends or 5'- or 3'-overhangs. Mot1-catalyzed dissociation of TBP from DNA was not prevented by a psoralen cross-link positioned immediately preceding the TATA sequence. Thus, Mot1 most likely promotes release of TBP from TATA-containing DNA by causing a structural change in TBP itself, rather than by strand unwinding.},

keywords = {Adenosine Triphosphatases, Base Sequence, Chromatography, DNA Helicases, DNA-Binding Proteins, Fungal, Gel, Gene Expression Regulation, Genetic, Kinetics, Molecular Sequence Data, Molecular Weight, Osmolar Concentration, Recombinant Proteins, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, TATA Box, TATA-Binding Protein Associated Factors, TATA-Box Binding Protein, Team-Mueller, Transcription, Transcription Factors},

pubstate = {published},

tppubtype = {article}

}

@article{,

title = {In vitro DNA and dGMP adducts formation caused by ochratoxin A},

author = { S. Obrecht-Pflumio and G. Dirheimer},

year  = {2000},

date = {2000-01-01},

journal = {Chem Biol Interact},

volume = {127},

number = {1},

pages = {29-44},

abstract = {Ochratoxin A (OTA), a nephrotoxic and nephrocarcinogenic mycotoxin, leads to the formation of DNA adducts after administration to animals. This could be due to an epigenetic effect. In vitro assays can exclude an indirect effect, where the xenobiotic can generate, in vivo, endogenous reactive compounds which give adducts on DNA. Microsomes prepared from mice or rabbit kidney and liver, used as metabolic activators, were incubated in the presence of commercial salmon testes DNA and OTA, with NADPH or arachidonic acid used as cofactors. Upto 126 DNA adducts for 10(9) nucleotides were detected using the 32P postlabeling method after incubation with the mouse kidney system. Similar results were obtained with rabbit kidney microsomes. Using liver microsomes, the number of DNA adducts detected was much lower. When NADPH was used as a cosubstrate (to explore the cytochrome P450 metabolic pathways), with mice kidney microsomes, the adduct level was only 44% of the one obtained with arachidonic acid. These results lend support to the hypothesis of the preferential activation of OTA by the peroxidase activity of prostaglandin synthases and/or lipoxygenases to direct genotoxic metabolites, and are in agreement with the previously obtained results after in vivo treatment of mice. In order to identify the nucleotides of DNA modified by the OTA metabolites, dAMP, dGMP, dTMP and dCMP were used as substrates under the same conditions as with DNA. The adducts were found only on dGMP. The total adduct level was of 344 adducts per 10(9) nucleotides with the appearance of three major adducts in the presence of arachidonic acid. With NADPH, 271 adducts were obtained per 10(9) nucleotides, with again three major adducts, but only two of them were similar to two adducts obtained in the presence of arachidonic acid. Desferal (desferrioxamine B methanesulphonate), at a 50 microM concentration, did not reduce the adduct level. Adducts were also obtained when polydG, polydC and dG-p-dG were used as alternative substrates, whereas no adducts were obtained with polydA, polydT and polydC. The major adduct obtained after incubation of DNA with OTA, comigrated with the major adduct obtained with dGMP, in two chromatographic solvents. These results show that OTA is metabolized to genotoxic metabolite(s) which interact with the guanine residues of DNA.},

note = {0009-2797 

Journal Article},

keywords = {Acid/metabolism, Adducts/*metabolism, Agents/pharmacology, Animals, Arachidonic, Carcinogens/pharmacology, Chelating, Chromatography, Deferoxamine/pharmacology, Deoxyguanine, DNA, Female, Kidney/ultrastructure, Liver/metabolism, Male, Mice, Microsomes, Microsomes/metabolism, Mycotoxins/pharmacology, NADP/metabolism, Nucleotides/*metabolism, Nucleotides/metabolism, Ochratoxins/*pharmacology, Peroxidase/metabolism, Rabbits, Spectrophotometry},

pubstate = {published},

tppubtype = {article}

}

@article{,

title = {Expression of an active form of recombinant Ty1 reverse transcriptase in Escherichia coli: a fusion protein containing the C-terminal region of the Ty1 integrase linked to the reverse transcriptase-RNase H domain exhibits polymerase and RNase H activities},

author = { M. Wilhelm and M. Boutabout and F. X. Wilhelm},

year  = {2000},

date = {2000-01-01},

journal = {Biochem J},

volume = {348},

number = {Pt 2},

pages = {337-42},

abstract = {Replication of the Saccharomyces cerevisiae Ty1 retrotransposon requires a reverse transcriptase capable of synthesizing Ty1 DNA. The first description of an active form of a recombinant Ty1 enzyme with polymerase and RNase H activities is reported here. The Ty1 enzyme was expressed as a hexahistidine-tagged fusion protein in Escherichia coli to facilitate purification of the recombinant protein by metal-chelate chromatography. Catalytic activity of the recombinant protein was detected only when amino acid residues encoded by the integrase gene were added to the N-terminus of the reverse transcriptase-RNase H domain. This suggests that the integrase domain could play a role in proper folding of reverse transcriptase. Several biochemical properties of the Ty1 enzyme were analysed, including the effect of MgCl(2), NaCl, temperature and of the chain terminator dideoxy GTP on its polymerase activity. RNase H activity was examined by monitoring the cleavage of a RNA-DNA template-primer. Our results suggest that the distance between the RNase H and polymerase active sites corresponds to the length of a 14-nucleotide RNA-DNA heteroduplex. The recombinant protein produced in E. coli should be useful for further biochemical and structural analyses and for a better understanding of the role of integrase in the activation of reverse transcriptase.},

note = {0264-6021 

Journal Article},

keywords = {&, Acid, affinity, Alignment, Amino, Calf, cerevisiae/*enzymology/*genetics, Chromatography, Cloning, Codon, coli, Comparative, Data, DNA, DNA/metabolism, Escherichia, Frames, Fusion, Genetic, Gov't, H, Heteroduplexes/metabolism, HIV-1, Homology, Integrases/chemistry/metabolism, Kinetics, Molecular, Non-U.S., Nucleic, Open, Polymerase/chemistry/isolation, Proteins/chemistry/isolation, purification/*metabolism, purification/metabolism, Reading, Recombinant, Retroelements/*genetics, Reverse, Ribonuclease, RNA-Directed, RNA/metabolism, Saccharomyces, Sequence, Study, Support, Templates, Terminator, Thymus/isolation, Transcriptase/chemistry},

pubstate = {published},

tppubtype = {article}

}

@article{lamberty_insect_1999,

title = {Insect immunity. Isolation from the lepidopteran Heliothis virescens of a novel insect defensin with potent antifungal activity},

author = {M Lamberty and S Ades and S Uttenweiler-Joseph and G Brookhart and D Bushey and Jules A Hoffmann and Philippe Bulet},

issn = {0021-9258},

year  = {1999},

date = {1999-04-01},

journal = {J. Biol. Chem.},

volume = {274},

number = {14},

pages = {9320--9326},

abstract = {Lepidoptera have been reported to produce several antibacterial peptides in response to septic injury. However, in marked contrast to other insect groups, no inducible antifungal molecules had been described so far in this insect order. Surprisingly, also cysteine-rich antimicrobial peptides, which predominate in the antimicrobial defense of other insects, had not been discovered in Lepidoptera. Here we report the isolation from the hemolymph of immune induced larvae of the lepidopteran Heliothis virescens of a cysteine-rich molecule with exclusive antifungal activity. We have fully characterized this antifungal molecule, which has significant homology with the insect defensins, a large family of antibacterial peptides directed against Gram-positive strains. Interestingly, the novel peptide shows also similarities with the antifungal peptide drosomycin from Drosophila. Thus, Lepidoptera appear to have built their humoral immune response against bacteria on cecropins and attacins. In addition, we report that Lepidoptera have conferred antifungal properties to the well conserved structure of antibacterial insect defensins through amino acid replacements.},

keywords = {Amino Acid, Animals, Antifungal Agents, Capillary, Chromatography, Defensins, Electrophoresis, Escherichia coli, Hemolymph, High Pressure Liquid, hoffmann, Insect Proteins, Larva, Lepidoptera, M3i, Micrococcus luteus, Proteins, Sequence Homology},

pubstate = {published},

tppubtype = {article}

}

@article{uttenweiler-joseph_differential_1998,

title = {Differential display of peptides induced during the immune response of Drosophila: a matrix-assisted laser desorption ionization time-of-flight mass spectrometry study},

author = {S Uttenweiler-Joseph and M Moniatte and Marie Lagueux and Van A Dorsselaer and Jules A Hoffmann and Philippe Bulet},

issn = {0027-8424},

year  = {1998},

date = {1998-09-01},

journal = {Proc. Natl. Acad. Sci. U.S.A.},

volume = {95},

number = {19},

pages = {11342--11347},

abstract = {We have developed an approach based on a differential mass spectrometric analysis to detect molecules induced during the immune response of Drosophila, regardless of their biological activities. For this, we have applied directly matrix-assisted laser desorption/ionization MS to hemolymph samples from individual flies before and after an immune challenge. This method provided precise information on the molecular masses of immune-induced molecules and allowed the detection, in the molecular range of 1.5-11 kDa, of 24 Drosophila immune-induced molecules (DIMs). These molecules are all peptides, and four correspond to already characterized antimicrobial peptides. We have further analyzed the induction of the various peptides by immune challenge in wild-type flies and in mutants with a compromised antimicrobial response. We also describe a methodology combining matrix-assisted laser desorption ionization time-of-flight MS, HPLC, and Edman degradation, which yielded the peptide sequence of three of the DIMs. Finally, molecular cloning and Northern blot analyses revealed that one of the DIMs is produced as a prepropeptide and is inducible on a bacterial challenge.},

keywords = {Animals, bacteria, Chromatography, Cloning, Hemolymph, High Pressure Liquid, hoffmann, Immunity, Insect Proteins, M3i, Mass, Matrix-Assisted Laser Desorption-Ionization, messenger, Molecular, Peptides, Protein Precursors, RNA, Sequence Analysis, Spectrometry, Time Factors},

pubstate = {published},

tppubtype = {article}

}

@article{taguchi_novel_1998,

title = {A novel insect defensin from the ant Formica rufa},

author = {S Taguchi and Philippe Bulet and Jules A Hoffmann},

issn = {0300-9084},

year  = {1998},

date = {1998-04-01},

journal = {Biochimie},

volume = {80},

number = {4},

pages = {343--346},

abstract = {By combination of size exclusion and reversed-phase chromatography, we have isolated a novel member of insect defensin-type antimicrobial peptides from the entire bodies of bacteria-challenged Formica rufa (hymenoptera, formicidae). The molecular mass of the purified peptide was estimated to be 4120.42 by matrix-assisted laser desorption/ionization-time of flight/mass spectrometry. Sequence analysis revealed that this peptide consisted of 40 amino acid residues with six cysteines engaged in the formation of three intramolecular disulfide bridges. This peptide is unique among the arthropod defensins in terms of the presence of asparatic acid and alanine at position 33 and as C-terminal residue, respectively. In addition, this novel defensin from Formica rufa has the particularity to have no C-terminal extension in contrast to those reported for other hymenoptera defensins.},

keywords = {Amino Acid, Animals, Anti-Bacterial Agents, Ants, Chromatography, High Pressure Liquid, hoffmann, Insect Proteins, insects, M3i, Mass, Matrix-Assisted Laser Desorption-Ionization, Protein Structure, Secondary, Sequence Alignment, Sequence Homology, Spectrometry},

pubstate = {published},

tppubtype = {article}

}

@article{ehret-sabatier_characterization_1996,

title = {Characterization of novel cysteine-rich antimicrobial peptides from scorpion blood},

author = {L Ehret-Sabatier and D Loew and M Goyffon and P Fehlbaum and Jules A Hoffmann and A van Dorsselaer and Philippe Bulet},

issn = {0021-9258},

year  = {1996},

date = {1996-11-01},

journal = {J. Biol. Chem.},

volume = {271},

number = {47},

pages = {29537--29544},

abstract = {We have isolated, from the hemolymph of unchallenged scorpions of the species Androctonus australis, three distinct antimicrobial peptides, which we have fully characterized by Edman degradation, electrospray ionization mass spectrometry, and matrix-assisted laser desorption/ionization mass spectrometry. Two are novel molecules: (i) androctonin, a 25-residue peptide with two disulfide bridges, active against both bacteria (Gram-positive and Gram-negative) and fungi and showing marked sequence homology to tachyplesins and polyphemusins from horseshoe crabs; and (ii) buthinin, a 34-residue antibacterial (Gram-positive and Gram-negative) peptide with three disulfide bridges. The third peptide contains 37 residues and three disulfide bridges and clearly belongs to the family of anti-Gram-positive insect defensins. We have synthesized androctonin and explored its activity spectrum and mode of action.},

keywords = {Animals, Anti-Bacterial Agents, Chromatography, Cysteine, Electron, Hemolymph, Hemolysis, High Pressure Liquid, hoffmann, M3i, Mass Spectrometry, Microscopy, Peptides, Scorpions},

pubstate = {published},

tppubtype = {article}

}

@article{charlet_innate_1996,

title = {Innate immunity. Isolation of several cysteine-rich antimicrobial peptides from the blood of a mollusc, Mytilus edulis},

author = {Maurice Charlet and S Chernysh and H Philippe and Charles Hetru and Jules A Hoffmann and Philippe Bulet},

issn = {0021-9258},

year  = {1996},

date = {1996-09-01},

journal = {J. Biol. Chem.},

volume = {271},

number = {36},

pages = {21808--21813},

abstract = {We have isolated from the blood of immune-challenged and untreated mussels (Mytilus edulis) antibacterial and antifungal peptides. We have characterized two isoforms of a novel 34-residue, cysteine-rich, peptide with potent bactericidal activity and partially characterized a novel 6.2-kDa antifungal peptide containing 12 cysteines. We report the presence of two members of the insect defensin family of antibacterial peptides and provide a phylogenetic analysis that indicates that mollusc and arthropod defensins have a common ancestry. Our data argue that circulating antimicrobial peptides represent an ancient host defense mechanism that predated the separation between molluscs and arthropods at the root of the Cambrian, about 545 million years ago.},

keywords = {Amino Acid, Animals, Anti-Infective Agents, Antifungal Agents, Bivalvia, Blood Proteins, Chromatography, Cysteine, Defensins, High Pressure Liquid, hoffmann, M3i, Molecular Weight, Phylogeny, Sequence Homology},

pubstate = {published},

tppubtype = {article}

}

@article{cociancich_novel_1994,

title = {Novel inducible antibacterial peptides from a hemipteran insect, the sap-sucking bug Pyrrhocoris apterus},

author = {S Cociancich and A Dupont and G Hegy and R Lanot and F Holder and Charles Hetru and Jules A Hoffmann and Philippe Bulet},

issn = {0264-6021},

year  = {1994},

date = {1994-06-01},

journal = {Biochem. J.},

volume = {300 ( Pt 2)},

pages = {567--575},

abstract = {Insects belonging to the recent orders of the endopterygote clade (Lepidoptera, Diptera, Hymenoptera and Coleoptera) respond to bacterial challenge by the rapid and transient synthesis of a battery of potent antibacterial peptides which are secreted into their haemolymph. Here we present the first report on inducible antibacterial molecules in the sap-sucking bug Pyrrhocoris apterus, a representative species of the Hemiptera, which predated the Endoptergotes by at least 50 million years in evolution. We have isolated and characterized from immune blood of this species three novel peptides or polypeptides: (i) a 43-residue cysteine-rich anti-(Gram-positive bacteria) peptide which is a new member of the family of insect defensins; (ii) a 20-residue proline-rich peptide carrying an O-glycosylated substitution (N-acetylgalactosamine), active against Gram-negative bacteria; (iii) a 133-residue glycine-rich polypeptide also active against Gram-negative bacteria. The proline-rich peptide shows high sequence similarities with drosocin, an O-glycosylated antibacterial peptide from Drosophila, and also with the N-terminal domain of diptericin, an inducible 9 kDa antibacterial peptide from members of the order Diptera, whereas the glycine-rich peptide has similarities with the glycine-rich domain of diptericin. We discuss the evolutionary aspects of these findings.},

keywords = {Amino Acid, Animals, Anti-Bacterial Agents, Antimicrobial Cationic Peptides, Blood Proteins, Chromatography, Defensins, Gas Chromatography-Mass Spectrometry, Gel, Gram-Negative Bacteria, Gram-Positive Bacteria, Hemiptera, Hemolymph, hoffmann, Insect Proteins, M3i, Peptides, Sequence Homology},

pubstate = {published},

tppubtype = {article}

}

@article{,

title = {Transfer RNA binding protein in the nucleus of Saccharomyces cerevisiae},

author = { M. L. Wilhelm and J. Reinbolt and J. Gangloff and G. Dirheimer and F. X. Wilhelm},

year  = {1994},

date = {1994-01-01},

journal = {FEBS Lett},

volume = {349},

number = {2},

pages = {260-4},

abstract = {A yeast nuclear protein that binds to tRNA was identified using a RNA mobility shift assay. Northwestern blotting and N-terminal sequencing experiments indicate that this tRNA-binding protein is identical to zuotin which has previously been shown to bind to Z-DNA [(1992) EMBO J. 11, 3787-3796]. Labeled tRNA and poly(dG-m5dC) stabilized in the Z-DNA form identify the same protein on a Northwestern blot. In a gel retardation assay poly(dG-m5dC) in the Z-form strongly diminishes the binding of tRNA to zuotin. These studies establish that zuotin is able to bind to both tRNA and Z-DNA. Zuotin may be transiently associated with tRNA in the nucleus of yeast cells and play a role in its processing or transport to the cytoplasm.},

note = {0014-5793 

Journal Article},

keywords = {*Saccharomyces, &, Acid, Amino, Cell, cerevisiae, cerevisiae/*metabolism, Chromatography, Data, DNA-Binding, DNA/metabolism, Fungal, Fungal/*isolation, high, liquid, Molecular, Nucleus/*metabolism, Pressure, Proteins, Proteins/genetics/*metabolism, purification, RNA, Saccharomyces, Sequence, Transfer/*isolation},

pubstate = {published},

tppubtype = {article}

}