Citation

Torres-Bonilla KA, Andrade-Silva D, Serrano SMT, Hyslop S. Comp. Biochem. Physiol. C Toxicol. Pharmacol. 2018; 213: 27-38.

Affiliation

Department of Pharmacology, Faculty of Medical Sciences, State University of Campinas (UNICAMP), Rua Tessália Vieira de Camargo, 126, Cidade Universitária Zeferino Vaz, 13083-887 Campinas, SP, Brazil. Electronic address: hyslop@fcm.unicamp.br.

Copyright

(Copyright © 2018, Elsevier Publishing)

DOI

10.1016/j.cbpc.2018.06.003

PMID

29966733

Abstract

In this work, we examined the proteolytic and phospholipase A₂ (PLA₂) activities of venom from the opisthoglyphous colubrid Pseudoboa neuwiedii. Proteolytic activity (3 and 10 μg of venom) was comparable to that of Bothrops neuwiedii venom but less than Bothrops atrox. This activity was inhibited by EDTA and 1,10-phenanthroline but only slightly affected (≤30% inhibition) by PMSF and AEBSF, indicating it was mediated by snake venom metalloproteinases (SVMPs). The pH and temperature optima for proteolytic activity were 8.0 and 37 °C, respectively. The venom had no esterase activity, whereas PLA₂ activity was similar to B. atrox, greater than B. neuwiedii but less than B. jararacussu. SDS-PAGE revealed venom proteins >100 kDa, 45-70 kDa, 21-24 kDa and ~15 kDa, and mass spectrometry of protein bands revealed SVMPs, cysteine-rich secretory proteins (CRISPs) and PLA₂, but no serine proteinases. In gelatin zymography, the most active bands occurred at 65-68 kDa (seen with 0.05-0.25 μg of venom). Caseinolytic activity occurred at 50-66 kDa and was generally weaker than gelatinolytic activity. RP-HPLC of venom yielded 15 peaks, five of which showed gelatinolytic activity; peak 7 was the most active and apparently contained a P-III class SVMP. The venom showed α-fibrinogenase activity, without affecting the β and γ chains; this activity was inhibited by EDTA and 1,10-phenanthroline. The venom did not clot rat citrated plasma but reduced the rate and extent of coagulation after plasma recalcification. In conclusion, P. neuwiedii venom is highly proteolytic and could potentially affect coagulation in vivo by degrading fibrinogen via SVMPs.

Copyright © 2018. Published by Elsevier Inc.

Language: en

Keywords

Back-fanged colubrid; Caseinolytic; Esterase; Fibrinogenase; Phospholipase A(2); Proteolytic; Pseudoboa neuwiedii; Snake venom metalloproteinase (SVMP)