Concanavalin A-Binding Enzymes of Crotalus scutulatus scutulatus Venom

Crotalus scutulatus scutulatus crude venom was separated into two fractions by Concanavalin A Sepharose 4B affinity chromatography. The proteins binding to Con A exhibited phosphomonoesterase (orthophosphoric monoester phosphohydrolase EC 3.1.3.2), phosphodiesterase, 5\u27-nucleotidase (5\u27-ribonucleotide phosphohydrolase EC 3.1.3.5), phospholipase A(phosphatidate 2-acylhydrolase EC 3.1.1 .4), hyaluronidase (hyaluronate glycanohydrolase EC 3.2.1 d), N-benzoyl-L-arginine ethyl esterase, p-toluenesulfonyl-L-arginine methyl esterase, L-amino acid oxidase (L-amino acid: 02 oxidoreductase [deaminating] EC 1.4.3.2), and caseinolytic activities. Thrombin-like and NAD nucleosidase (5\u27-ribonucleotide phosphohydrolase EC 3.1.3.5) activities were not observed. The crude venom and the fraction containing the glycoproteins which bound to Con A were fractionated by DEAE Sephadex A-50 ion exchange chromatography. Each of these samples yielded fractions having caseinolytic activities

Concanavalin A-Nonbinding Enzymes of Crotalus scutulatus scutulatus Venom

Crotalus scutulatus scutulatus crude venom was separated into two fractions by Concanavalin A Sepharose 4B affinity chromatography. The Concanavalin A-nonbinding fraction (F-l) exhibited phosphomonoesterase (orthophosphoric monoester phosphohydrolase EC 3.1 .3.2), phosphodiesterase, 5 \u27-nucleotidase (5 \u27-ribonucleotide phosphohydrolase EC 3.1.3.5), phospholipase A (phosphatidate 2-acylhydrolase EC 3.1.1.4), hyaluronidase (hyaluronate glycanohydrolase EC 3.2.1.d), N-benzoyl-Larginine ethyl esterase, p-toluenesulfonyl-L-arginine methyl esterase, L-amino acid oxidase (L-amino acid: O2 oxidoreductase [deaminating] EC 1.4.3.2), and caseinolytic activities. Thrombin-like and NAD nucleosidase (5 \u27-ribonudeotide phosphohydrolase EC 3.1.3.5) activities were not observed. DEAE Sephadex A-50 ion exchange chromatography by two stage elution of F-l yielded several fractions having proteinase activities. Proteinase activity was observed in the latter fractions of the first elution and in the fractions of the second elution

Enzymes in Heloderma horridum Venom

A mixture of venom and saliva from the lizard Heloderma horridum was analyzed for esterase, phosphomonoesterase, phosphodiesterase, 5\u27nucleotidase, and protease activities. Hydrolysis of N-benzoyl-L-arginine ethyl ester occurred at a pH optimum between pH 8.6 and 9.1 with a maximum activity of 452 units per mg per min. Hydrolysis of ptoluenesulfonyl-L-arginine methyl ester occurred at a pH optimum between pH 8.1 and 8.5 with a maximum of only 36 units per mg per min. One mg of the venom mixture liberated 9.3 μM of p nitrophenol from p-nitrophenyl phosphate per minute at an optimum pH between 8.2 and 8.3. Over a wide range of pH, only low phosphodiesterase and 5\u27nucleotidase activities were observed. A trace of caesinolytic activity occurred at pH 9.0

5\u27-Nucleotidase and Thrombin-Like Activities of Selected Crotalid Venoms

Thrombin-like activities were not observed inCrotalus basiliscus, C. molossus and C. scutulatus scutulatus crude venoms. 5\u27-Nucleotidase specific activities of 0.863, 0.273 and 5.520 units/mg of crude venom protein were observed inC. basiliscus, C. molossus and C. s. scutulatus venoms, respectively. Concanavalin ASepharose 4 B (Con A)affinitychromatography yielded two fractions from each of the crude venoms. Ineach instance, both fractions exhibited 5\u27-nucleotidase activities and the Con A-binding proteins had higher activities than the Con A-nonbinding proteins. 5\u27-Nucleotidase activities inthe DEAESephadex A-50 chromatographic fractions were localized in the first elution fraction and the last fraction(s) to elute. EDTAhad no effect on the 5\u27-nucleotidase activities ofthe crude venoms