PURIFICATION AND PARTIAL CHARACTERIZATION OF A THROMBIN-LIKE GYROXIN ENZYME FROM BUSHMASTER (LACHESIS-MUTA-RHOMBEATA) VENOM/

A. S. Aguiar, C. R. Alves, A. R. Melgarejo and S. Giovanni-De-Simone. Purification and partial characterization of a thrombin-like/gyroxin e nzyme from bushmaster (Lachesis muta rhombeata) venom. Toxicon 34, 555 -565, 1996.-The acidic coagulating enzyme of the L. m. rhombeata venom was purified to homogeneity using one step on preparative isoelectric focusing followed by gel permeation on a high performance liquid chro matography system. The enzyme focused with pIs 3.1-5.0 and had a molec ular mass of 47,000 mel. wt as determined by high performance liquid g el-filtration chromatography and about 45,000 mel. wt as judged by sod ium dodecyl sulfate-polyacrylamide-gel electrophoresis. The enzyme is a glycoprotein containing sialic acid and 12.4% of neutral carbohydrat es. The 30 N-terminal amino acid sequence of the L. m. rhombeata prote in shows 100% identity with L. m. muta gyroxin and considerable sequen ce homology with gyroxin and thrombin-related proteins. The enzyme exh ibits strong N-p-tosyl-L-arginine methyl esterase activity, hydrolyses tripeptide nitroanilide derivatives weakly or not at all, and cleaves specifically the fibropeptide A (alpha-chain). Copyright (C) 1996 Els evier Science Ltd.