Format

Send to

Choose Destination
J Mass Spectrom. 2007 Nov;42(11):1405-14.

Snake venomics. Strategy and applications.

Author information

1
Instituto de Biomedicina de Valencia, CSIC, Jaume Roig 11, 46010 Valencia, Spain. jcalvete@ibv.csic.es

Abstract

Snake bites can be deadly, but the venoms also contain components of medical and biotechnological value. The proteomic characterization of snake venom proteomes, snake venomics, has thus a number of potential benefits for basic research, clinical diagnosis, and development of new research tools and drugs of potential clinical use. Snake venomics is also relevant for a deep understanding of the evolution and the biological effects of the venoms, and to generate immunization protocols to elicit toxin-specific antibodies with greater specificity and effectiveness than conventional systems. Our snake venomics approach starts with the fractionation of the crude venom by reverse-phase HPLC, followed by the initial characterization of each protein fraction by combination of N-terminal sequencing, SDS-PAGE, and mass spectrometric determination of the molecular masses and the cysteine (SH and S--S) content. Protein fractions showing a single electrophoretic band, molecular mass, and N-terminal sequence can be straightforwardly assigned by BLAST analysis to a known protein family. On the other hand, protein fractions showing heterogeneous or blocked N-termini are analyzed by SDS-PAGE and the bands of interest subjected to automated reduction, carbamidomethylation, and in-gel tryptic digestion. The resulting tryptic peptides are then analyzed by MALDI-TOF mass fingerprinting followed by amino acid sequence determination of selected doubly and triply charged peptide ions by collision-induced dissociation tandem mass spectrometry. The combined strategy allows us to assign unambiguously all the isolated venom toxins representing over 0.05% of the total venom proteins to known protein families. Protocols and applications of snake venomics are reviewed and discussed.

PMID:
17621391
DOI:
10.1002/jms.1242
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Wiley
Loading ...
Support Center