RT Journal Article
SR Electronic
T1 Purine Nucleoside Phosphorylase: Kinetics, Mechanism, and Specificity
JF Molecular Pharmacology
JO Mol Pharmacol
FD American Society for Pharmacology and Experimental Therapeutics
SP 526
OP 536
VO 3
IS 6
A1 KRENITSKY, THOMAS A.
YR 1967
UL http://molpharm.aspetjournals.org/content/3/6/526.abstract
AB Ribosyl exchange reactions between purines and purine nucleosides catalyzed by crystalline purine nucleoside phosphorylase from calf spleen were markedly stimulated by inorganic phosphate. Initial velocity and product inhibition analyses were performed for the enzymic synthesis of inosine from ribose 1-phosphate and hypoxanthine. An ordered sequential mechanism involving the isomerization of time free enzyme and the initial combination of the purine base with one of the forms of time free enzyme is most consistent with these results. The specificity of the enzyme toward the purine base in the synthesis of ribonucleosides was investigated. In order of decreasing effectiveness, ribosyl acceptors were guanine, hypoxanthine, xanthine, 6-mercaptopurine, and allopurinol [4-hydroxypyrazolo(3,4-d)pyrimidine]. Oxoallopurinol [4,6-dihydroxypyrazolo-(3,4-d)pyrimidine] was a very poor substrate. Adenine, azathioprine [6-(1-mlethyl-4-nitro-5-imidazolyl)thiopurine], and uracil did not produce detectable amounts of ribonucleoside. Guanosine, deoxyguanosine, and inosine were shown to be effective pentosyl donors in exchange reactions. ACKNOWLEDGMENTS To G. B. Elion and G. H. Hitchings, the authors gratitude for their help and advice in the conduct of these studies and the preparation of the manuscript.