A heat stable paraoxonase (O,O-diethyl O-p-nitrophenyl phosphate O-p-nitrophenyl hydrolase) from Russell's viper venom.
Fractionation of Russell's viper venom revealed separate phosphohydrolase activities directed against p-nitrophenyl phosphate, bis(p-nitrophenyl) phosphate, p-nitrophenylthymidylic acid, and O,O-diethyl p-nitrophenyl phosphate (paraoxon). On gel fractionation, the first two activities are eluted ahead of the latter. They could be resolved further by phosphocellulose cation exchange chromatography. The hydrolytic activities directed against p-nitrophenylthymidylic acid hydrolyzing component is heat labile, while the paraoxon hydrolyzing component manifests an unusually high degree of heat stability. Gel filtration yields 9600 for the molecular weight of the "paraoxonase". This enzyme, as all known enzymes of this type, requires the presence of a divalent cation. Maximum activity is obtained in the presence of Ca2+. In the presence of Sr2+ the reaction rate is 50% of that of Ca2+; other divalent cations show lower activities. The presence of the enzyme is species specific. Of four species tested, only Russell's viper venom showed significant paraoxonase activity. Enzyme activity is intact following incubation with iodoacetate of p-chloromercuribenzoate. Activity is partially preserved even in the presence of 8 M urea.
D009597:4-Nitrophenylphosphatase / Q000302:isolation & purification / Q000378:metabolism; D000818:Animals; D002413:Cations, Divalent; D004355:Drug Stability; D004950:Esterases / Q000302:isolation & purification* / Q000378:metabolism; D006358:Hot Temperature; D007700:Kinetics; D010261:Paraoxon; D012910:Snake Venoms / Q000032:analysis*; D012911:Snakes; D013329:Structure-Activity Relationship
D002413:Cations, Divalent; D012910:Snake Venoms; D004950:Esterases; D009597:4-Nitrophenylphosphatase; D010261:Paraoxon