Uracil phosphoribosyltransferase from the extreme thermoacidophilic archaebacterium Sulfolobus shibatae is an allosteric enzyme, activated by GTP and inhibited by CTP.
Uracil phosphoribosyltransferase, which catalyses the formation of UMP and pyrophosphate from uracil and 5-phosphoribosyl alpha-1-pyrophosphate (PRPP), was partly purified from the extreme thermophilic archaebacterium Sulfolobus shibatae. The enzyme required divalent metal ions for activity and it showed the highest activity at pH 6.4. The specific activity of the enzyme was 50-times higher at 95 degrees C than at 37 degrees C, but the functional half-life was short at 95 degrees C. The activity of uracil phosphoribosyltransferase was strongly activated by GTP, which increased Vmax of the reaction by approximately 20-fold without much effect on K(m) for the substrates. The concentration of GTP required for half-maximal activation was about 80 microM. CTP was a strong inhibitor and acted by raising the concentration of GTP needed for half-maximal activation of the enzyme. We conclude that uracil phosphoribosyltransferase from S. shibatae is an allosteric enzyme which is activated by a purine nucleotide and inhibited by a pyrimidine nucleotide as seen for several enzymes in the pyrimidine nucleotide biosynthetic pathway of Escherichia coli, but not observed before for any phosphoribosyltransferase.
Linde
L
L
Center for Enzyme Research, Institute of Molecular Biology, University of Copenhagen, Denmark.
Jensen
K F
KF
D000255:Adenosine Triphosphate / Q000378:metabolism / Q000494:pharmacology; D000494:Allosteric Regulation; D003570:Cytidine Triphosphate / Q000378:metabolism* / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D006160:Guanosine Triphosphate / Q000378:metabolism* / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D010430:Pentosyltransferases / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism*; D010754:Phosphoribosyl Pyrophosphate / Q000378:metabolism; D013379:Substrate Specificity; D016944:Sulfolobus / Q000737:chemistry / Q000201:enzymology*; D013696:Temperature
D003570:Cytidine Triphosphate; D010754:Phosphoribosyl Pyrophosphate; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D010430:Pentosyltransferases; C023269:uracil phosphoribosyltransferase