[en] The crude venom of Conus virgo was analyzed by Fourier transform mass spectrometry (FTMS) using both nano-electrospray ionization and MALDI. The analyses were performed directly on the crude venom, without chromatographic separation. The mass fingerprinting of the venom yielded 64 distinct molecular masses in the range 500-4500 Da with two major components at 1328.5142 and 1358.5592 Da. To facilitate the de novo sequencing of these compounds, the disulfide bonds of all components were reduced for the whole venom. The mass accuracy, resolution and sensitivity provided by FTMS were necessary to complete the sequencing of the two new peptides named ViVA and ViVB, that turned out to be conotoxins belonging to the T-superfamily, with the disulfide framework V. The peptides shared 80% similarity and as often observed for this class of compound, they were highly post-translationally modified: amidated C-terminus, pyroglutamic acid residue at the N-terminus and two disulfide bonds. Complementary online nano-LC-nano-ESI-FTMS experiments were undertaken. Among the 130 molecular masses found in the coupling experiments, only 45 were common with those obtained in the direct approach, which means that 21 compounds observed by nano-ESI-FTMS were not detected. This clearly shows that some discriminations against some classes of compounds occur when a chromatographic step is used before mass spectrometry.
Disciplines :
Chemistry
Author, co-author :
Quinton, Loïc ; Ecole Polytechnique (France) > Laboratoire des Mécanismes Réactionnels - Palaiseau- France
Le Caër, Jean-Pierre; Ecole Polytechnique (France) > Laboratoire des Mécanismes Réactionnels - Palaiseau- France
Vinh, Joëlle; Ecole Supérieure de Physique et de Chimie Industrielles de la ville de Paris > France
Gilles, Nicolas; Commissariat à l'Energie Atomique (Saclay) - CEA > Département d'ingénierie et d'Etude des Proteines - Gif-sur-Yvette- France
Chamot-Rooke, Julia; Ecole Polytechnique (France) > Laboratoire des Mécanismes Réactionnels - Palaiseau- France
Language :
English
Title :
Fourier transform mass spectrometry: A powerful tool for toxin analysis
Publication date :
May 2006
Journal title :
Toxicon
ISSN :
0041-0101
eISSN :
1879-3150
Publisher :
Pergamon Press - An Imprint of Elsevier Science, Oxford, United Kingdom
Bao Y., Bu P., Jin L., Hongxia W., Yang Q., and An L. Purification, characterization and gene cloning of a novel phospholipase A2 from the venom of Agkistrodon blomhoffii ussurensis. Int. J. Biochem. Cell Biol. 37 (2005) 558-565
Breci L.A., Tabb D.L., Yates III J.R., and Wysocki V.H. Cleavage N-terminal to proline: analysis of a database of peptide tandem mass spectra. Anal. Chem. 75 (2003) 1963-1971
Burmeister Getz E.L., Xiao M., Chakrabarty T., Cooke R., and Selvin P.R. A comparison between the sulfhydryl reductants tris(2-carboxyethyl)phosphine and dithiothreitol for use in protein biochemistry. Anal. Chem. 273 (1999) 73-80
Burn J.A., Butler J.C., Moran J., and Whitesides G.M. Selective reduction of disulfides by tris(2-carboxyethyl)phosphine. J. Org. Chem. 56 (1991) 2648-2650
Caravatti P., and Allemann M. The infinity cell-a new trapped-ion cell with radiofrequency covered trapping electrodes for fourier-transform ion-cyclotron resonance mass-spectrometry. Org. Mass Spectrom. 26 (1991) 514-518
Cooper H.J., Hakansson K., and Marshall A.G. The role of electron capture dissociation in biomolecular analysis. Mass Spectrom. Rev. 24 (2005) 201-222
Diego-García E., Batista C.V.F., García-Gómez B.I., Lucas S., Candido D.M., Gómez-Lagunas F., and Possani L.D. The Brazilian scorpion Tityus costatus Karsch: genes, peptides and function. Toxicon 45 (2005) 273-283
Dwoskin L.P., and Crooks P.A. Competitive neuronal nicotinic receptor antagonists: a new direction for drug discovery. J. Pharmacol. Exp. Ther. 298 (2001) 395-402
Gauthier J.W., Trautman T.R., and Jacobson D.B. Sustained off-resonance irradiation for collision-activated dissociation involving fourier transform mass spectrometry. Collision-activated dissociation technique that emulates infrared multiphoton dissociation. Anal. Chem. Acta 246 (1991) 211-225
Gorman J.J., Wallis T.P., and Pitt J.J. Protein disulfide bond determination by mass spectrometry. Mass Spectrom. Rev. 21 (2002) 183-216
Hansson K., Furie B., Furie B.C., and Stenflo J. Isolation and characterization of three novel Gla-containing conus marmoreus venom peptides, one with a novel cystein pattern. Biochem. Biophys. Res. Commun. 319 (2004) 1081-1087
Juraschek R., Dulcks T., and Karas M. Nanoelectrospray-more than just a minimized-flow electrospray ionization source. J. Am. Soc. Mass Spectrom. 10 (1999) 300-308
Kalume D.E., Stenflo J., Czerwiec E., Hambe B., Furie B.C., Furie B., and Roepstorff P. Structure determination of two conotoxins from conus textile by a combination of matrix-assisted laser desorption/ionization time-of-flight and electrospray mass spectrometry and biochemical methods. J. Mass Spectrom. 35 (2000) 145-156
Kelleher N.L., Zubarev R.A., Bush K., Furie B.C., Mc Lafferty F.W., and Walsh C.T. Localization of labile posttranslational modifications by electron capture dissociation: the case of gamma-carboxyglutamic acid. Anal. Chem. 71 (1999) 4250-4253
Li L., Masselon C.D., Anderson G.A., Pasa-Tolic L., Lee S.-W., Shen Y., Zhao R., Lipton M.S., Conrads T.P., Tolic N., and Smith R.D. High-throughput peptide identification from protein digests using data-dependent multiplexed tandem FTICR mass spectrometry coupled with capillary liquid chromatography. Anal. Chem. 73 (2001) 3312-3322
Liska A.J., and Shevchenko A. Combining mass spectrometry with database interrogation strategies in proteomics. Trends Anal. Chem. 22 (2003) 291-298
Marshall A.G. Accurate mass measurement: taking full advantage of nature's istotopic complexity. Physica B 346-347 (2004) 503-508
Marshall A.G., and Hendrickson C.L. Fourier transform ion cyclotron resonance detection: principles and experimental configurations. Int. J. Mass Spectrom. 215 (2002) 59-75
Marshall A.G., Hendrickson C.L., and Jackson G.S. Fourier transform ion cyclotron resonance mass spectrometry: a primer. Mass Spectrom. Rev. 17 (1998) 1-35
Olivera B.M. Conus venom peptides, receptor and ion channel targets, and drug design: 50 million years of neuropharmacology. Mol. Biol. Cell 8 (1997) 2101-2109
Olivera B.M., Rivier J., Clark C., Ramilo C.A., Corpuz G.P., Abogadie F.C., Mena E.E., Woodward S.R., Hillyard D.R., and Cruz L.J. Diversity of Conus neuropeptides. Science 249 (1990) 257-263
Quinton, L., Le Caer, J.-P., Phan, G., Ligny-Lemaire, C., Bourdais-Jomaron, J., Ducancel, F., Chamot-Rooke, J., 2005. Characterization of new toxins within crude venoms by combined use of fourier transform mass spectrometry and cloning. Anal. Chem. 77, 6630-6639.
Roepstorff P., and Fohlman J. Proposal for a common nomenclature for sequence ions in mass spectra of peptides. Biomed. Mass Spectrom. 11 (1984) 601
Schlosser A., and Lehmann W.D. Patchwork peptide sequencing: extraction of sequence information from accurate mass data of peptide tandem mass spectra recorded at high resolution. Proteomics 2 (2002) 524-533
Sharpe I.A., Gehrmann J., Loughnan M.L., Thomas L., Adams D.A., Atkins A., Palant E., Craik D.J., Adams D.J., Alewood P.F., and Lewis R.J. Two new classes of conopeptides inhibit the α-1-adrenoreceptor and noradrenaline transporter. Nat. Neurosci. 4 (2001) 902-907
Spengler B. De novo sequencing, peptide composition analysis, and composition-based sequencing: a new strategy employing accurate mass determination by fourier transform ion cyclotron resonance mass spectrometry. J. Am. Soc. Mass Spectrom. 15 (2004) 704-715
Taggi A.E., Meinwald J., and Schroeder F.C. A new approach to natural products discovery exemplified by the identification of sulfated nucleosides in spider venom. J. Am. Chem. Soc. 126 (2004) 10364-10369
Tzouros M., Manov N., Bienz S., and Bigler L. Tandem mass spectrometric investigation of acylpolyamines of spider venoms and their 15N-labeled derivatives. J. Am. Soc. Mass Spectrom. 11 (2004) 1636-1643
Walker C.S., Steel D., Jacobsen R.B., Lirazan M.B., Cruz L.J., Hooper D., Shetty R., DelaCruz R.C., Nielsen J.S., Zhou L.M., Bandyopadhyay P., Craig A.G., and Olivera B.M. The T-superfamily of conotoxins. J. Biol. Chem. 274 (1999) 30664-30671
Wermelinger L.S., Dutra D.L.S., Oliveira-Carvalho A.L., Soraes M.R., Bloch Jr. C., and Zingali R.B. Fast analysis of low molecular mass compounds present in snake venoms: identification of ten new pyroglutamate-containing peptides. Rapid Commun. Mass Spectrom. 19 (2005) 1703-1708
Wilm M., and Mann M. Analytical properties of the nanoelectrospray source. Anal. Chem. 68 (1996) 1-9
Wysocki V.H., Tsaprailis G., Smith L.L., and Breci L.A. Mobile and localized protons: a framework for understanding peptide dissociation. J. Mass Spectrom. 35 (2000) 1399-1406
Yates III J.R. Mass spectrometry: from genomics to proteomics. Trends Genet. 16 (2000) 5-8
