Metabolism of purine nucleosides in human and ovine lymphocytes and rat thymocytes and their influence on mitogenic stimulation.
1. Phosphorolysis and phosphorylation rates of inosine, guanosine and deoxyguanosine were determined in disrupted and intact human and ovine lymphocytes and rat thymocytes and related with their effect on mitogenic stimulation. 2. Activity of purine nucleoside phosphorylase (EC 2.4.2.1) was about 10 times higher in extracts of human lymphocytes than in those of ovine lymphocytes and rat thymocytes. Apparent Km values for inosine and guanosine were higher in human lymphocytes (about 100 microM) than in ovine lymphocytes (50 microM). Apparent Km values for deoxyguanosine were about 100 microM in the extracts of all three cell types. 3. In extracts of human and ovine lymphocytes the presence of guanosine kinase activity was established. Deoxyguanosine kinase activity was detected in all three cell types. 4. The rate of phosphorylation of deoxyguanosine was much lower than the rate of phosphorolysis both in extracts and in intact cells. 5. Deoxyguanosine, guanosine and inosine were incorporated by intact cells into nucleotides and nucleic acids. This incorporation of deoxyguanosine and guanosine was only partially due to phosphorolysis and subsequent conversion by hypoxanthine-guanine phosphoribosyltransferase (EC 2.4.2.8). The incorporation of inosine appeared to be due completely to this route. 6. Inosine (0.5 mM) did not inhibit thymidine incorporation of phytohemagglutinin-stimulated human and ovine lymphocytes. At the same concentration deoxyinosine caused 50% inhibition, but guanosine and deoxyguanosine inhibited almost completely. Thymidine incorporation of concanavalin A-stimulated rat thymocytes was hardly inhibited by 0.5 mM inosine, deoxyinosine and guanosine, but 50 microM and 0.5 mM deoxyguanosine caused 25% and complete inhibition, respectively.
D000818:Animals; D003208:Concanavalin A; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000378:metabolism; D007700:Kinetics; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000276:immunology / Q000378:metabolism*; D008297:Male; D010766:Phosphorylation; D011684:Purine Nucleosides / Q000378:metabolism*; D011685:Purine Nucleotides / Q000378:metabolism; D011683:Purine-Nucleoside Phosphorylase / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D012756:Sheep; D013045:Species Specificity; D013950:Thymus Gland / Q000378:metabolism
D011684:Purine Nucleosides; D011685:Purine Nucleotides; D003208:Concanavalin A; D011683:Purine-Nucleoside Phosphorylase; D007041:Hypoxanthine Phosphoribosyltransferase