CAMPERI Silvia Andrea
congresos y reuniones científicas
One-Bead-One-Peptide Library to Purify Crotalus durissus terrificus Phosholipase A2
M. C. MARTÍNEZ-CERON; S. L. SAAVEDRA; L. ÁVILA; S. L. GIUDICESSI; F. ALBERICIO; S. A. CAMPERI; O. CASCONE
Simposio; American Peptide Symposium.; 2015
Crotalus durissus terrificus (Cdt), is the only species of that genus in Argentina. Crotoxin represents more than 50% of the dry weight of its venom. This protein has two subunits: A (phospholipase A2, PLA2), and B (crotapotin). Venom profile differ according to geographical regions. The aim of this work was to purify Argentinean Cdt PLA2 to analyze its characteristics and possible uses as antiviral activity against tropical diseases. Short peptides are stable and resistant to proteases and can be produced in high quantities and purity. In this work, a one-bead-one-peptide library (XXXXXGGAGG; X= A, E, F, H, L, N, P, R, S, T, V, Y) was synthesized using the Divide-Couple-Recombine method (DCR). Screening was done using pure PLA2 labeled with NHS-Biotin. Those peptides with affinity for the protein were revealed using Streptavidin-Peroxidase and α-Chloronaphtol/H2O2. They were isolated to release the peptides from HMBA-ChemMatrix using ammonia vapor. Peptides were analyzed by MALDI-TOF MSMS. Fifty peptides were sequenced and their amino acid moieties were studied. Two of them were selected to be immobilized in Sepharose. Both peptides were capable to purify PLA2 in one step with high yield.