pyrophosphate and purine

1. The formation of adenosine 5'-phosphate, guanosine 5'-phosphate and inosine 5'-phosphate from [8-(14)C]adenine, [8-(14)C]guanine and [8-(14)C]hypoxanthine respectively in the presence of 5-phosphoribosyl pyrophosphate and an extract from Ehrlich ascites-tumour cells was assayed by a method involving liquid-scintillation counting of the radioactive nucleotides on diethylaminoethylcellulose paper. The results obtained with guanine were confirmed by a spectrophotometric assay which was also used to assay the conversion of 6-mercaptopurine and 5-phosphoribosyl pyrophosphate into 6-thioinosine 5'-phosphate in the presence of 6-mercaptopurine phosphoribosyltransferase from these cells. 2. At pH 7.8 and 25 degrees the Michaelis constants for adenine, guanine and hypoxanthine were 0.9 mum, 2.9 mum and 11.0 mum in the assay with radioactive purines; the Michaelis constant for guanine in the spectrophotometric assay was 2.6 mum. At pH 7.9 the Michaelis constant for 6-mercaptopurine was 10.9 mum. 3. 25 mum-6-Mercaptopurine did not inhibit adenine phosphoribosyltransferase. 6-Mercaptopurine is a competitive inhibitor of guanine phosphoribosyltransferase (K(i) 4.7 mum) and hypoxanthine phosphoribosyltransferase (K(i) 8.3 mum). Hypoxanthine is a competitive inhibitor of guanine phosphoribosyltransferase (K(i) 3.4 mum). 4. Differences in kinetic parameters and in the distribution of phosphoribosyltransferase activities after electrophoresis in starch gel indicate that different enzymes are involved in the conversion of adenine, guanine and hypoxanthine into their nucleotides. 5. From the low values of K(i) for 6-mercaptopurine, and from published evidence that ascites-tumour cells require supplies of purines from the host tissues, it is likely that inhibition of hypoxanthine and guanine phosphoribosyltransferases by free 6-mercaptopurine is involved in the biological activity of this drug.

Topics: Adenine; Adenine Nucleotides; Adenosine Monophosphate; Animals; Ascites; Carcinoma, Ehrlich Tumor; Diphosphates; Electrophoresis; Enzyme Inhibitors; Glucosyltransferases; Guanine; Guanine Nucleotides; Hypoxanthines; Kinetics; Mercaptopurine; Nucleotides; Pharmacology; Purines; Research; Spectrophotometry; Transferases; Xanthines

Topics: Adenine; Adenine Nucleotides; Animals; Carbon Isotopes; Chromatography; Cytosine; Cytosine Nucleotides; Diphosphates; Fishes; Guanine; Guanine Nucleotides; Metabolism; Nucleotides; Orotic Acid; Phosphoribosyl Pyrophosphate; Purines; Pyrimidines; Research; Salmon; Thymine; Tissue Extracts; Tritium; Uracil; Uracil Nucleotides; Xanthines

Topics: Adenine Nucleotides; Antimetabolites; Antineoplastic Agents; Azaguanine; Chromatography; Diphosphates; Enzyme Inhibitors; Glutamine; Mercaptopurine; Nucleotides; Purines; Pyrazoles; Pyrimidines; Research; Thioguanine; Transferases; Ultraviolet Rays

Topics: Adenine Nucleotides; Alcohols; Chemical Phenomena; Chemistry, Physical; Dinucleoside Phosphates; Diphosphates; Methanol; Polynucleotides; Purines; Research; Solvents; Spectrophotometry; Ultraviolet Rays; Urea

Topics: Adenine Nucleotides; Adenosine Triphosphate; Amides; Amidophosphoribosyltransferase; Birds; Diphosphates; Enzyme Inhibitors; Feedback; Glutamine; Guanine Nucleotides; Hypoxanthines; Liver; Purines; Research; Ribonucleotides; Ribose; Transferases

Topics: Adenine; Animals; Bone Marrow; Chick Embryo; Diphosphates; DNA; In Vitro Techniques; Liver; Metabolism; Pentosephosphates; Pharmacology; Phosphoribosyl Pyrophosphate; Purines; Rabbits; Rats; Research; RNA; Thymine

Topics: Amidophosphoribosyltransferase; Diphosphates; Humans; Nucleosides; Nucleotides; Purines; Transferases