Phosphoribosylpyrophosphate synthesis in cultured human cells

Phosphoribosylpyrophosphate (PRPP) concentrations and PRPP synthetase activity were studied in cultured human fibroblasts with control and mutant hypoxanthine-guanine phosphoribosylpyrophosphate (HPRT) activity. The PRPP concentrations increased 20- to 50-fold and PRPP synthetase activity 3-fold in cells from patients with the Lesch-Nyham syndrome when aminopterin, an inhibitor of de novo purine synthesis, was added to the medium. Concentrations of PRPP and PRPP synthetase activity did not increase in control cells in medium containing aminopterin unless hypoxanthine was removed from the medium. Exposure of cells to cycloheximide, a protein synthesis inhibitor, prevented the induction of PRPP synthetase and the formation of high PRPP concentrations. Cells from a patient with a mutant HPRT with a high Michaelis constant for PRPP increased PRPP levels and PRPP synthetase activity to a new steady state when they were grown in medium containing aminopterin. Inhibitors of de novo purine synthesis, the presence of hypoxanthine in the medium, and mutant HPRT activity affect the regulation of PRPP levels and PRPP synthetase activity.