Reference : New method for characterizing highly disulfide-bridged peptides in complex mixtures: App...
Scientific journals : Article
Physical, chemical, mathematical & earth Sciences : Chemistry
http://hdl.handle.net/2268/1173
New method for characterizing highly disulfide-bridged peptides in complex mixtures: Application to toxin identification from crude venoms
English
Quinton, Loïc mailto [Université de Liège - ULg > Département de chimie (sciences) > Chimie physique, spectrométrie de masse >]
Demeure, Kevin [Université de Liège - ULg > Département de chimie (sciences) > Chimie physique, spectrométrie de masse >]
Dobson, Rowan mailto [Université de Liège - ULg > Département de chimie (sciences) > Chimie physique, spectrométrie de masse >]
Gilles, Nicolas [CEA/DSV/iBiTec-S/SIMOPRO, Gif-sur-Yvette Cedex 91191, France > > > >]
Gabelica, Valérie mailto [Université de Liège - ULg > Département de chimie (sciences) > Chimie physique, spectrométrie de masse >]
De Pauw, Edwin mailto [Université de Liège - ULg > Département de chimie (sciences) > Chimie physique, spectrométrie de masse >]
2007
Journal of Proteome Research
Amer Chemical Soc
6
8
3216-3223
Yes
International
1535-3893
Washington
[en] in-source decay ; post-source decay ; conotoxin ; snake toxins ; disulfide-bridged peptides ; top-down sequencing ; mass spectrometry ; venom
[en] Animal venoms are highly complex mixtures that can contain many disulfide-bridged toxins. This work presents an LC-MALDI approach allowing (1) a rapid classification of toxins according to their number of disulfide bonds and (2) a rapid top-down sequencing of the toxins using a new MALDI matrix enhancing in-source decay (ISD). The crude venom is separated twice by LC: the fractions of the first separation are spotted on the MALDI matrix alpha-cyano-4-hydroxycinnamic acid (CHCA) and the others using 1,5-diaminonaphthalene (1,5-DAN). CHCA spots are more convenient for obtaining a precise mass fingerprint of a large number of peptides; however, the analysis of 1,5-DAN spots allows the number of disulfide bridges to be counted owing to their partial in-plume reduction by this particular matrix. Subsequently, the disulfide bonds of all peptides present in the crude venom were reduced by an excess of tris(carboxyethyl)phosphine before the LC separation and were subjected to the same analysis in CHCA and 1,5-DAN. Toxins were sequenced using a TOF/TOF analysis of metastable fragments from CHCA spots and ISD fragmentation from 1,5-DAN spots. Novel conotoxin sequences were found using this approach. The use of 1,5-DAN for ISID top-down sequencing is also illustrated for higher molecular weight toxins such as snake cardiotoxins and neurotoxins (>6500 Da), where sequence coverage >70% is obtained from the c-ion series.
Giga-Systems Biology and Chemical Biology
Fonds de la Recherche Scientifique (Communauté française de Belgique) - F.R.S.-FNRS
Researchers
http://hdl.handle.net/2268/1173
10.1021/pr070142t

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