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(Reference retrieved automatically from SciELO through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Purification procedure for the isolation of a P-I metalloprotease and an acidic phospholipase A 2 fromBothrops atrox snake venom

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Author(s):
Danilo L. Menaldo [1] ; Anna L. Jacob-Ferreira [2] ; Carolina P. Bernardes [3] ; Adélia C. O. Cintra [4] ; Suely V. Sampaio [5]
Total Authors: 5
Affiliation:
[1] Universidade de São Paulo. Faculdade de Ciências Farmacêuticas de Ribeirão Preto. Departamento de Análises Clínicas, Toxicológicas e Bromatológicas - Brasil
[2] Universidade de São Paulo. Faculdade de Ciências Farmacêuticas de Ribeirão Preto. Departamento de Análises Clínicas, Toxicológicas e Bromatológicas - Brasil
[3] Universidade de São Paulo. Faculdade de Ciências Farmacêuticas de Ribeirão Preto. Departamento de Análises Clínicas, Toxicológicas e Bromatológicas - Brasil
[4] Universidade de São Paulo. Faculdade de Ciências Farmacêuticas de Ribeirão Preto. Departamento de Análises Clínicas, Toxicológicas e Bromatológicas - Brasil
[5] Universidade de São Paulo. Faculdade de Ciências Farmacêuticas de Ribeirão Preto. Departamento de Análises Clínicas, Toxicológicas e Bromatológicas - Brasil
Total Affiliations: 5
Document type: Journal article
Source: Journal of Venomous Animals and Toxins including Tropical Diseases; v. 21, p. 0-0, 2015-09-29.
Abstract

Background Snake venoms are complex mixtures of inorganic and organic components, mainly proteins and peptides. Standardization of methods for isolating bioactive molecules from snake venoms is extremely difficult due to the complex and highly variable composition of venoms, which can be influenced by factors such as age and geographic location of the specimen. Therefore, this study aimed to standardize a simple purification methodology for obtaining a P-I class metalloprotease (MP) and an acidic phospholipase A2 (PLA 2 ) from Bothrops atroxvenom, and biochemically characterize these molecules to enable future functional studies.Methods To obtain the toxins of interest, a method has been standardized using consecutive isolation steps. The purity level of the molecules was confirmed by RP-HPLC and SDS-PAGE. The enzymes were characterized by determining their molecular masses, isoelectric points, specific functional activity and partial amino acid sequencing.Results The metalloprotease presented molecular mass of 22.9 kDa and pI 7.4, with hemorrhagic and fibrin(ogen)olytic activities, and its partial amino acid sequence revealed high similarity with other P-I class metalloproteases. These results suggest that the isolated metalloprotease is Batroxase, a P-I metalloprotease previously described by our research group. The phospholipase A 2 showed molecular mass of 13.7 kDa and pI 6.5, with high phospholipase activity and similarity to other acidic PLA2 s from snake venoms. These data suggest that the acidic PLA2 is a novel enzyme from B. atrox venom, being denominated BatroxPLA 2 .Conclusions The present study successfully standardized a simple methodology to isolate the metalloprotease Batroxase and the acidic PLA 2 BatroxPLA2 from the venom of B. atrox, consisting mainly of classical chromatographic processes. These two enzymes will be used in future studies to evaluate their effects on the complement system and the inflammatory process, in addition to the thrombolytic potential of the metalloprotease. (AU)

FAPESP's process: 12/11963-1 - Effects of a metalloprotease and an acidic phospholipase A2 from Bothrops atrox snake venom on the complement system and the inflammatory process
Grantee:Danilo Luccas Menaldo
Support Opportunities: Scholarships in Brazil - Post-Doctoral
FAPESP's process: 11/23236-4 - Native and recombinant animal toxins: functional, structural and molecular analysis
Grantee:Suely Vilela
Support Opportunities: Research Projects - Thematic Grants
FAPESP's process: 12/21569-9 - ANTITHROMBOTIC AND TROMBOLYTIC ACTIVITIES OF BATROXASE, A METALLOPROTEINASE PURIFIED FROM THE VENOM OF Bothrops atrox SNAKE, IN EXPERIMENTAL ANIMAL MODELS.
Grantee:Anna Laura Bechara Jacob Ferreira
Support Opportunities: Scholarships in Brazil - Post-Doctoral