Some regulatory properties of purine biosynthesis de novo in long-term cultures of epithelial-like rat liver cells

De novo synthesis of purine nucleotides and some regulatory properties of this pathway were studied in cultured epithelial-like rat liver cells. It was found that the physiological 5-phosphoribosyl 1-pyrophosphate (P-Rib-PP) concentration in these cells is limiting for purine synthesis de novo. Increase of P-Rib-PP availability, achieved by activation of P-Rib-PP synthetase at high P_i concentration, resulted in acceleration of purine synthesis. The effects of increasing cellular ribose 5-phosphate (Rib-5-P availability, by methylene blue-induced acceleration of the oxidative pentose phosphate pathway, on P-Rib-PP availability and on the rate of the novo purine synthesis were also studied. It was found that at the P_i concentration prevailing in the tissue at extracellular physiological P_i concentration, Rib-5-P availability is saturating for P-Rib-PP generation and therefore also for purine synthesis.