The influence of ribose 5-phosphate availability on purine synthesis of cultured human lymphoblasts and mitogen-stimulated lymphocytes.
The intracellular ribose 5-phosphate concentration was found to be an important determinant of rates of de novo purine synthesis. When ribose 5-phosphate production was reduced in cultured human lymphoblasts by glucose starvation, the intracellular phosphoribosylpyrophosphate concentration and rates of de novo purine synthesis decreased. Inosinate-guanylate:pyrophosphate phosphoribosyltransferase (HPR transferase)-deficient cells were relatively more resistant to glucose starvation. To minimize the effect of purine nucleotide feedback inhibition on the de novo pathway, cells were treated with inhibitors of IMP dehydrogenase and adenylosuccinate synthetase. In normal lymphoblasts, purine synthesis was stimulated only at glucose concentrations greater than 100 microM while in HPR transferase-deficient lymphoblasts, stimulation occurred even in the absence of glucose. The differences between the normal and HPR transferase-deficient cells were lost when ribose reutilization from endogenous nucleotide breakdown was impaired in the HPR transferase-deficient cells by incubation with 2'-deoxyinosine. Endogenous ribose reutilization for purine synthesis is, therefore, important when either glucose availability is limited or synthesis is stimulated. In the absence of glucose, exogenous purine nucleotides restored the intracellular concentrations of ribose 5-phosphate, phosphoribosylpyrophosphate, and purine nucleotides to almost 100% and rates of purine synthesis to 50-75% of those at 10 mM glucose. When ribose 5-phosphate production was increased in peripheral blood lymphocytes by phytohemagglutinin activation, the intracellular phosphoribosylpyrophosphate concentration and rates of de novo purine synthesis increased.
D002460:Cell Line; D002478:Cells, Cultured; D006801:Humans; D007700:Kinetics; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000276:immunology / Q000378:metabolism*; D010428:Pentosephosphates / Q000378:metabolism*; D011685:Purine Nucleotides / Q000378:metabolism; D011687:Purines / Q000096:biosynthesis*; D012267:Ribosemonophosphates / Q000378:metabolism*
D010428:Pentosephosphates; D011685:Purine Nucleotides; D011687:Purines; D012267:Ribosemonophosphates; C031626:ribose-5-phosphate