Influence of substituent ribose on transition state affinity in reactions catalyzed by adenosine deaminase.
Adenosine deaminase from calf intestine hydrolyzes adenine at a limiting rate four orders of magnitude lower than that for adenosine, while Km values for these substrates are about the same (Wolfenden, R., et al. (1969), Biochemistry 8, 2412-2415). Reactivity of 6-substituents, toward nucleophilic displacement, is found to be affected only slightly by removal of ribose as a 9-substituent, in model reactions. Substituent ribose thus appears to stabilize, selectively, the transition state for enzymatic deamination. In contrast with the small influence of substituent ribose on the apparent binding affinity of substrates, removal of substituent ribose from a potential transition state analogue, 1,6-dihydro-6-hydroxy-methylpurine ribonucleoside, results in a lowering of its affinity for the enzyme by several orders of magnitude. The synthesis of the analogue and related compounds is described, and their properties compared with those of other photoadducts and of the naturally occurring inhibitors covidarabine and coformycin. Binding of these inhibitors is found to result in the appearance of ultraviolet-absorbing bands in the neighborhood of 323 nm.
D000243:Adenosine Deaminase / Q000378:metabolism; D000345:Affinity Labels; D000818:Animals; D002417:Cattle; D007422:Intestines / Q000201:enzymology; D007700:Kinetics; D009700:Nucleoside Deaminases / Q000378:metabolism; D012266:Ribose; D013056:Spectrophotometry, Ultraviolet; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
D000345:Affinity Labels; D012266:Ribose; D009700:Nucleoside Deaminases; D000243:Adenosine Deaminase