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Structural studies with snake venom phospholipases A2: native, recombinant and complexed with miotoxic activity inhibitors.

Grant number: 12/10830-8
Support type:Scholarships in Brazil - Doctorate
Effective date (Start): November 01, 2012
Effective date (End): February 29, 2016
Field of knowledge:Biological Sciences - Biophysics - Molecular Biophysics
Principal researcher:Marcos Roberto de Mattos Fontes
Grantee:Guilherme Henrique Marchi Salvador
Home Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil


Snakes from Bothrops genus represents 90% of ophidian accidents that occurs in Brazil. One of the components of snake venoms is phospholipases A2 (PLA2), which are small molecules and shows a diversity of biological activities. In the last years, several studies with PLA2s and homologue PLA2s from snake venoms in native and complexes forms where made to better understand of the action mechanism of these toxins. In this work, will be performed studies with PLA2s and homologue PLA2s from Bothrops moojeni snake venom native, complexed with inhibitors and PLAs homologue from Bothrops pirajai in recombinant form. For this, will be done: i) structural analysis of BmooPLA2-I, a PLA2 isolated from B. moojeni snake venom, ii) purification of MjTX-I and MjTX-II from the whole venom and co-crystallization with caffeic acid (CA) and rosmarinic acid (RA) and iii) production of recombinant PrTX-I and possible mutants, to proof the proposed Lys49-PLA2s myotoxic site functionality. The BmooPLA2-I has a hypotensive action and inhibit the platelet aggregation, such as BthA-I (PLA2 isolated from B. jararacussu snake venom). Recently the BmooPLA2 structure was elucidated in our lab and was observed several structural similarities to BthA-I. MjTX-I is a homologue PLA2 with the amino acids sequence and crystallographic structure very different of others Lys49-PLA2s; and in this project, we propose the co-crystallization of MjTX-I and MjTX-II with inhibitors CA and RA, which are compounds found in various plants and popularly known to have anti-ophidian properties. Studies of recombinants toxins will be performed with PrTX-I, since this protein can be considered a model for studies and this snake is currently in the endangered animals list.

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Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
SALVADOR, Guilherme Henrique Marchi. Structural studies of complex between phospholipases A2 homologues isolated from Bothrops moojeni venom and myotoxic activity inhibitors. 2016. Doctoral Thesis - Universidade Estadual Paulista (Unesp). Instituto de Biociências. Botucatu Botucatu.

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