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(Reference retrieved automatically from Web of Science 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) from Bothrops atrox snake venom

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Author(s):
Menaldo, Danilo L. [1] ; Jacob-Ferreira, Anna L. [1] ; Bernardes, Carolina P. [1] ; Cintra, Adelia C. O. [1] ; Sampaio, Suely V. [1]
Total Authors: 5
Affiliation:
[1] Univ Sao Paulo, Dept Anal Clin Toxicol & Bromatol, Fac Ciencias Farmaceut Ribeirao Preto, BR-14040903 Ribeirao Preto, SP - Brazil
Total Affiliations: 1
Document type: Journal article
Source: Journal of Venomous Animals and Toxins including Tropical Diseases; v. 21, AUG 13 2015.
Web of Science Citations: 8
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 A(2) (PLA(2)) from Bothrops atrox venom, 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 PLA(2)s from snake venoms. These data suggest that the acidic PLA(2) 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) BatroxPLA(2) 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: 12/21569-9 - 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
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: 12/11963-1 - 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