Phosphorolytic and ribosyl transfer mechanisms of purified chicken liver purine nucleoside phosphorylase.

Abstract

Purified chicken liver purine nucleoside phosphorylase shows two ionizable groups at the active site whose pKa were near pH 6.9 and 8; the molecular weight (67,000-89,000) depends on the protein concentration. Initial velocity studies and product inhibition patterns were consistent with a random mechanism, which is rapid equilibrium in the phosphorolytic reaction with a dead-end complex, but not in the synthetic reaction. Free inorganic orthophosphate purine nucleoside phosphorylase (Sephadex G-100) catalyzes a pentosyl transfer reaction from inosine to guanine according to a random Bi, Bi mechanism.