Enzymatic characterization of a novel phospholipase A2 from Crotalus durissus cascavella rattlesnake (maracambóia) venom

被引：25

作者：

Beghini D.G. ^{[1
]}

Toyama M.H. ^{[1
,2
]}

Hyslop S. ^{[1
,3
]}

Sodek L.C. ^{[2
]}

Novello ^{[1
]}

Marangoni S. ^{[1
]}

机构：

[1] Departamento de Bioquímica, Instituto de Biologia, Universidade Estadual de Campinas (UNICAMP), CEP 13083-970, Campinas, SP

[2] Departamento de Fisiologia, Instituto de Biologia (IB), Univesidade Estadual de Campinas (UNICAMP), Campinas, SP

[3] Departamento de Farmacologia, Faculdade de Ciências Médicas, Universidade Estadual de Campinas (UNICAMP), Campinas, SP

来源：

Journal of Protein Chemistry | 2000年 / 19卷 / 8期

关键词：

Crotalus durissus cascavella; Crotapotin; Crotoxin; PLA[!sub]2[!/sub; Rattlesnake; Venom;

D O I：

10.1023/A:1007152303179

中图分类号：

学科分类号：

摘要：

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.

引用

页码：679 / 684

页数：5

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