Biochemical characterization of venom from Pseudoboa neuwiedii (Neuwied's false boa; Xenodontinae; Pseudoboini)

Author(s)

Torres-Bonilla, Kristian A. ^[1] ; Andrade-Silva, Debora ^[2] ; Serrano, Solange M. T. ^[2] ; Hyslop, Stephen ^[1]

Affiliation

^[1] State Univ Campinas UNICAMP, Fac Med Sci, Dept Pharmacol, Rua Tessalia Vieira Camargo, 126, Cidade Univ, BR-13083887 Campinas, SP - Brazil

^[2] Inst Butantan, Ctr Toxins Immune Response & Cell Signaling CeTIC, Lab Especial Toxinol Aplicada, Ave Brazil 1500, Sao Paulo, SP - Brazil

• Document type: Journal article
• Source: COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY C-TOXICOLOGY & PHARMACOLOGY; v. 213, p. 27-38, NOV 2018.
• Web of Science Citations: 3

Abstract

In this work, we examined the proteolytic and phospholipase A(2) (PLA(2)) activities of venom from the opisthoglyphous colubrid Pseudoboa neuwiedii. Proteolytic activity (3 and 10 mu g 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 degrees C, respectively. The venom had no esterase activity, whereas PLA(2) 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 similar to 15 kDa, and mass spectrometry of protein bands revealed SVMPs, cysteine-rich secretory proteins (CRISPs) and PLA(2), but no serine proteinases. In gelatin zymography, the most active bands occurred at 65-68 kDa (seen with 0.05-0.25 mu g 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 alpha-fibrinogenase activity, without affecting the beta and gamma 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. (AU)

• FAPESP's process: 13/07467-1 - CeTICS - Center of Toxins, Immune-Response and Cell Signaling
• Grantee: Hugo Aguirre Armelin
• Support Opportunities: Research Grants - Research, Innovation and Dissemination Centers - RIDC