title: Purification and partial characterization of a thrombin-like/gyroxin enzyme from bushmaster (Lachesis muta rhombeata) venom.
abstract: 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 chromatography system. The enzyme focused with pIs 3.1-5.0 and had a molecular mass of 47,000 mol. wt as determined by high performance liquid gel-filtration chromatography and about 45,000 mol. wt as judged by sodium dodecyl sulfate-polyacrylamide-gel electrophoresis. The enzyme is a glycoprotein containing sialic acid and 12.4% of neutral carbohydrates. The 30 N-terminal amino acid sequence of the L. m. rhombeata protein shows 100% identity with L. m. muta gyroxin and considerable sequence homology with gyroxin and thrombin-related proteins. The enzyme exhibits 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).
authors.affiliation: Instituto Vital Brazil, Niterói RJ, Brazil.
authors.forename: A S
authors.initials: AS
authors.lastname: Aguiar
authors.forename: C R
authors.initials: CR
authors.lastname: Alves
authors.forename: A
authors.initials: A
authors.lastname: Melgarejo
authors.forename: S
authors.initials: S
authors.lastname: Giovanni-de-Simone
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D050260:Carbohydrate Metabolism; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D003435:Crotalid Venoms / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D006868:Hydrolysis; D007525:Isoelectric Focusing; D008969:Molecular Sequence Data; D008970:Molecular Weight; D019158:N-Acetylneuraminic Acid / Q000378:metabolism; D010447:Peptide Hydrolases / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013917:Thrombin / Q000737:chemistry; D014757:Viper Venoms / Q000201:enzymology / Q000302:isolation & purification*; D017819:Viperidae
chemical_list: D003435:Crotalid Venoms; D014757:Viper Venoms; C017356:gyroxin; D010447:Peptide Hydrolases; C023073:tosylarginine methyl ester hydrolase; D013917:Thrombin; D019158:N-Acetylneuraminic Acid