Citation

Petras D, Heiss P, Süssmuth RD, Calvete JJ. J. Proteome Res. 2015; 14(6): 2539-2556.

Copyright

(Copyright © 2015, American Chemical Society)

DOI

10.1021/acs.jproteome.5b00305

PMID

25896403

Abstract

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 eleven three-finger toxins (7-7.9 kDa), a Kunitz-type inhibitor (6.3 kDa), ohanin (11.9 kDa), a novel phospholipase A2 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 complete 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 through Southeast Asia. Its venom composition, comprising 32-35 proteins/peptides from 10 protein families, is dominated by α-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 components, and locus-resolved toxin identification through top-down and bottom-up MS/MS-based species-specific database searching and de novo sequencing, promises a bright future to the field of venom research.

Language: en