Purification and characterisation of chicken brain hypoxanthine-guanine phosphoribosyltransferase.

Hypoxanthine-guanine phosphoribosyltransferase enzyme (EC 2.4.2.8) from chicken brain has been purified 10 000-fold to homogeneity. The molecular mass of the native enzyme is 85 kDa, with four subunits, each of 26 kDa, and exerts its maximum activity at pH 10.0. The Km values for hypoxanthine and guanine are 5.2 and 1.8 microM, respectively. The half-life of the enzyme is 30 min at 85 degrees C. Monoclonal antibodies were raised against the native purified enzyme and were used for purification of enzyme to homogeneity. The monoclonal antibody did not bind to the active centre of the enzyme.