Journal
JOURNAL OF PROTEOME RESEARCH
Volume 14, Issue 6, Pages 2539-2556Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.jproteome.5b00305
Keywords
Snake venomics; Elapidae; king cobra; Ophiophagus hannah; top-down proteomics; top-down venomics
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Funding
- Deutsche Forschungsgemeinschaft, Cluster of Excellence Unifying Concepts in Catalysis (UniCat)
- Ministerio de Ciencia e Innovacion, Madrid [BFU2010-17373]
- former Ministerio de Economia y Competitividad, Madrid [BFU2013-42833-P]
- CYTED project [BIOTOX P211RT0412]
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We report on the first application of top-down mass spectrometry in snake venomics. De novo sequence tags generated by, and ProSight Lite supported analysis of, combined collisional based dissotiations (CID and HCD) recorded in a hybrid LTQ Orbitrap instrument in data-dependent mode identified a number of proteins from different toxin families, namely, 11 three-finger toxins (7-7.9 kDa), a Kunitz-type inhibitor (6.3 kDa), ohanin (11.9 kDa), a novel phospholipase A(2) molecule (13.8 kDa), and the cysteine-rich secretory protein (CRISP) ophanin (25 kDa) from Indonesian king cobra venom. Complementary bottom-up MS/MS analyses contributed to the completion of a locus-resolved venom phenotypic map for Ophiophagus hannah, the world's longest venomous snake and a species of medical concern across its wide distribution range in forests from India to Southeast Asia. Its venom composition, comprising 32-35 proteins/peptides from 10 protein families, is dominated by alpha-neurotoxins and convincingly explains the main neurotoxic effects of human envenoming caused by king cobra bite. The integration of efficient chromatographic separation of the venom's components and locus-resolved toxin identification through top-down and bottom-up MS/MS-based species-specific database searching and de novo sequencing holds promise that the future will be bright for the field of venom research.
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