Selective expression of phosphoribosylpyrophosphate synthetase superactivity in human lymphoblast lines.
Phenotypic expression of 5-phosphoribosyl 1-pyrophosphate (PRPP) synthetase superactivity was examined in lymphoblast lines derived from six unrelated male patients. Fibroblasts from these individuals have increased rates of PRPP and purine nucleotide synthesis and express four classes of kinetic derangement underlying enzyme superactivity: increased maximal reaction velocity (catalytic defect); inhibitor resistance (regulatory defect); increased substrate affinity (substrate binding defect); and combined catalytic and regulatory defects. Lymphoblast lines from three patients with catalytic defects and from three normal individuals were indistinguishable with respect to enzyme activities, PRPP concentrations and generation, and rates of purine synthesis. Enzyme in lymphoblasts from a patient with combined defects also showed normal maximal reaction velocity but expressed purine nucleotide inhibitor resistance. A second regulatory defect and a substrate binding defect were also demonstrable in lymphoblasts and were identical to the enzyme defects in fibroblasts from the respective patients. Regulatory and substrate binding defects in lymphoblasts were accompanied by increased rates of PRPP and purine nucleotide synthesis. Among explanations for selective expression of enzyme superactivity, reduced concentrations of catalytically superactive enzymes seemed unlikely: immunoreactive PRPP synthetase was comparable in normal-derived and patient-derived cells. Activation of normal enzyme in transformed lymphocytes was also unlikely because absolute specific activities of lymphoblast PRPP synthetases corresponded to those of normal fibroblast and erythrocyte enzymes. Abnormal electrophoretic mobilities and thermal stabilities, identified in certain catalytically superactive fibroblast PRPP synthetases, were not found in the corresponding lymphoblast enzymes. Thus, lymphoblast PRPP synthetases from patients with catalytic superactivity appeared to differ structurally and functionally from their fibroblast counterparts.
M J
MJ
Losman
D
D
Rimon
M
M
Kim
M A
MA
Becker
D000494:Allosteric Regulation; D002384:Catalysis; D002460:Cell Line; D005347:Fibroblasts / Q000201:enzymology; D006412:Hematopoietic Stem Cells / Q000201:enzymology*; D006801:Humans; D008214:Lymphocytes / Q000201:enzymology; D008297:Male; D010754:Phosphoribosyl Pyrophosphate / Q000096:biosynthesis; D010770:Phosphotransferases / Q000096:biosynthesis*; D011687:Purines / Q000096:biosynthesis; D012268:Ribose-Phosphate Pyrophosphokinase / Q000096:biosynthesis*; D013379:Substrate Specificity
D011687:Purines; D010754:Phosphoribosyl Pyrophosphate; D010770:Phosphotransferases; D012268:Ribose-Phosphate Pyrophosphokinase