Enzymic characterization of Bacillus subtilis GTP cyclohydrolase I. Evidence for a chemical dephosphorylation of dihydroneopterin triphosphate.
GTP cyclohydrolase I catalyses the first committing step in the biosynthesis of the pterin moiety of folic acid: conversion of GTP to dihydroneopterin triphosphate. GTP cyclohydrolase I of Bacillus subtilis was purified to homogeneity and shown to have a homo-octameric structure. The enzyme had an apparent Km for GTP of 4 microM and, in the absence of cations, a Vmax. of 80 nmol/min per mg of protein. K+ ions moderately increased its Vmax., whereas UTP and Ca2+ and Mg2+ ions drastically increased its Km for GTP. Dihydrofolate and other products of the folate and tetrahydrobiopterin pathways did not inhibit GTP cyclohydrolase I. In addition to their effect on the enzyme activity, Ca2+ and Mg2+ ions catalysed the chemical dephosphorylation of dihydroneopterin triphosphate to non-cyclic dihydroneopterin monophosphate, the substrate for the phosphomonoesterase reaction in folate biosynthesis. This dephosphorylation was specific and did not require the action of a phosphatase. We suggest a physiological role for Ca2+ ions and UTP in regulation of folate biosynthesis at the levels of GTP cyclohydrolase I and dephosphorylation of dihydroneopterin triphosphate.
De Saizieu
A
A
Biotechnology Section, F. Hoffmann-La Roche Ltd., Basel, Switzerland.
Vankan
P
P
van Loon
A P
AP
D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics; D001483:Base Sequence; D001667:Binding, Competitive; D002118:Calcium / Q000494:pharmacology; D005492:Folic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D006136:GTP Cyclohydrolase / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D006160:Guanosine Triphosphate / Q000378:metabolism; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D008969:Molecular Sequence Data; D019798:Neopterin / Q000031:analogs & derivatives; D009876:Operon; D010766:Phosphorylation; D010957:Plasmids; D011188:Potassium / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D011621:Pteridines / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D014544:Uridine Triphosphate / Q000494:pharmacology
D011621:Pteridines; D011994:Recombinant Proteins; C013490:dihydroneopterin triphosphate; C010920:dihydrofolate; D019798:Neopterin; D006160:Guanosine Triphosphate; D005492:Folic Acid; D006136:GTP Cyclohydrolase; D008274:Magnesium; D011188:Potassium; D002118:Calcium; D014544:Uridine Triphosphate