Stereochemistry of hydrolysis by snake venom phosphodiesterase.
[18O]Adenosine 5'-O-phosphorothioate-O-p-nitrophenyl ester was prepared by saponification of the bis (-O,O-p-nitrophenyl ester) with K18OH. Only the diastereoisomer with the Rp configuration si a substrate for snake venom phosphodiesterase. The asymmetrically labeled [18O]adenosine 5'-O-phosphorothioate formed in this reaction was converted enzymatically to [18O]adenosine 5'-(1-thiodiphosphate) with the Sp configuration. The position of the 18O label, either bridging [1,2-mu-18O] or nonbridging [1-18O] was then determined. The results show that the reaction catalyzed by snake venom phosphodiesterase takes place with retention of configuration at phosphorus. This indicates that the hydrolysis proceeds via a covalent nucleotide enzyme intermediate.
D003435:Crotalid Venoms; D006868:Hydrolysis; D009218:Myosins; D010103:Oxygen Isotopes; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism*; D013379:Substrate Specificity
D003435:Crotalid Venoms; D010103:Oxygen Isotopes; D010727:Phosphoric Diester Hydrolases; D009218:Myosins