caa a22 4500 475744357 CHVBK 20180406123508.0 cr unu---uuuuu 170329e20001101xx s 000 0 eng 10.1023/A:1007152303179 doi (NATIONALLICENCE)springer-10.1023/A:1007152303179 Enzymatic Characterization of a Novel Phospholipase A2 from Crotalus durissus cascavella Rattlesnake (Maracambóia) Venom [Elektronische Daten] [D. Beghini, M. Toyama, S. Hyslop, L. Sodek, Novello,, S. Marangoni] The PLA2 and crotapotin subunits of crotoxin from Crotalus durissus cascavella venom were purified by a combination of high-performance liquid chromatography (HPLC) molecular exclusion (Protein Pack 300SW column) and reverse-phase HPLC (RP-HPLC). Tricine SDS-PAGE showed that the PLA2 and crotapotins migrated as single bands with estimated molecular masses of 15 and 9 kDa, respectively. The amino acid composition of the PLA2 showed the presence of 14 half-cysteines and a high content of basic residues (Lys, Arg, His), whereas the crotapotins were rich in hydrophobic, negatively charged residues and half-cysteines. The PLA2 showed allosteric behavior, with maximal activity at pH 8.3 and 35-40°C. C. d. cascavella PLA2 required Ca2+ for activity but was inhibited by Cu2+ and Zn2+ and by Cu2+ and Mg2+ in the presence and absence of Ca2+, respectively. Crotapotin (F3) and heparin inhibited the catalytic activity of the PLA2 by acting as allosteric inhibitors. Plenum Publishing Corporation, 2000 Crotalus durissus cascavella nationallicence crotoxin nationallicence crotapotin nationallicence PLA2 nationallicence rattlesnake nationallicence venom nationallicence Beghini D. Departamento de Bioquímica, Instituto de Biologia, Universidade Estadual de Campinas (UNICAMP), CP 6109, CEP 13083-970, Campinas, SP, Brazil aut Toyama M. Departamento de Bioquímica, Instituto de Biologia, Universidade Estadual de Campinas (UNICAMP), CP 6109, CEP 13083-970, Campinas, SP, Brazil aut Hyslop S. Departamento de Farmacologia, Faculdade de Ciências Médicas, Universidade Estadual de Campinas (UNICAMP), Campinas, SP, Brazil aut Sodek L. Departamento de Fisiologia, Instituto de Biologia (IB), (, Univesidade Estadual de Campinas (UNICAMP), Campinas, SP, Brazil aut Novello, Departamento de Bioquímica, Instituto de Biologia, Universidade Estadual de Campinas (UNICAMP), CP 6109, CEP 13083-970, Campinas, SP, Brazil aut Marangoni S. Departamento de Bioquímica, Instituto de Biologia, Universidade Estadual de Campinas (UNICAMP), CP 6109, CEP 13083-970, Campinas, SP, Brazil aut Journal of Protein Chemistry Kluwer Academic Publishers-Plenum Publishers 19/8(2000-11-01), 679-684 0277-8033 19:8<679 2000 19 10930 https://doi.org/10.1023/A:1007152303179 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1007152303179 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Beghini D. Departamento de Bioquímica, Instituto de Biologia, Universidade Estadual de Campinas (UNICAMP), CP 6109, CEP 13083-970, Campinas, SP, Brazil aut NATIONALLICENCE 700 1- Toyama M. Departamento de Bioquímica, Instituto de Biologia, Universidade Estadual de Campinas (UNICAMP), CP 6109, CEP 13083-970, Campinas, SP, Brazil aut NATIONALLICENCE 700 1- Hyslop S. Departamento de Farmacologia, Faculdade de Ciências Médicas, Universidade Estadual de Campinas (UNICAMP), Campinas, SP, Brazil aut NATIONALLICENCE 700 1- Sodek L. Departamento de Fisiologia, Instituto de Biologia (IB), (, Univesidade Estadual de Campinas (UNICAMP), Campinas, SP, Brazil aut NATIONALLICENCE 700 1- Novello Departamento de Bioquímica, Instituto de Biologia, Universidade Estadual de Campinas (UNICAMP), CP 6109, CEP 13083-970, Campinas, SP, Brazil aut NATIONALLICENCE 700 1- Marangoni S. Departamento de Bioquímica, Instituto de Biologia, Universidade Estadual de Campinas (UNICAMP), CP 6109, CEP 13083-970, Campinas, SP, Brazil aut NATIONALLICENCE 773 0- Journal of Protein Chemistry Kluwer Academic Publishers-Plenum Publishers 19/8(2000-11-01), 679-684 0277-8033 19:8<679 2000 19 10930 Metadata rights reserved Springer special CC-BY-NC licence nationallicence SWISSBIB 475744136 BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer