Crystallization and X-ray diffraction data collection of Basic PLA2 from Crotalus durrissus terrificus
L. C. Corrêa1, D. P. Marchi-Salvador1, C. Z. Oliveira2, A. M. Soares2 and M.R.M. Fontes1
1Dept. de Física e Biofísica, Instituto de Biociências, UNESP, Botucatu-SP; fontes@ibb.unesp.br 2Dept. de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, USP-SP
Phospholipases A2 are small stable calcium-dependent enzymes which belong to the superfamily of proteins that hydrolyzes the sn-2 ester bond of sn-3 membrane phospholipids to release arachidonic acid and lysophospholipids. These proteins are involved in several biochemical reactions and biological effects including neurotoxicity, myotoxicity, edema formatives, hypotension and anticoagulant action.
Basic PLA2 is present in crotalic venoms forming a protein complex with crotapotin increasing its neurotoxic potency. Recent studies showed that it also presents antimicrobial activity. Basic PLA2 is an Asp49-PLA2 consisting of 121 amino acids (Mr= 14,7 kDa). Here, we report the crystallization and X- ray diffraction data collection results of this new protein. Crystals were obtained by hanging drop method, with the condition: 0.1 M Tris HCl pH 9,0 and 11% PEG 8000 at 291K after twenty days. X- ray diffraction data of a single basic PLA2 crystal were collected at 100K using a Syncrotron Radiation Source (LNLS, Campinas, Brazil). Data were processed using Denzo/ Scalepack program at 2,28 Å resolution. The crystals belong to P212121 space group with cell constants a= 72.895, b= 81.159, c= 100.039Å. There are four monomers in the assimetric unit according with Matthews coefficient 2,642 Å3/Da and solvent fraction 51%.
This work has received financial supported from FAPESP, CNPq and LNLS.
|