Histidine 179 mutants of GTP cyclohydrolase I catalyze the formation of 2-amino-5-formylamino-6-ribofuranosylamino-4(3H)-pyrimidinone triphosphate.
GTP cyclohydrolase I catalyzes the conversion of GTP to dihydroneopterin triphosphate. The replacement of histidine 179 by other amino acids affords mutant enzymes that do not catalyze the formation of dihydroneopterin triphosphate. However, some of these mutant proteins catalyze the conversion of GTP to 2-amino-5-formylamino-6-ribofuranosylamino-4(3H)-pyrimidinone 5'-triphosphate as shown by multinuclear NMR analysis. The equilibrium constant for the reversible conversion of GTP to the ring-opened derivative is approximately 0.1. The wild-type enzyme converts the formylamino pyrimidine derivative to dihydroneopterin triphosphate; the rate is similar to that observed with GTP as substrate. The data support the conclusion that the formylamino pyrimidine derivative is an intermediate in the overall reaction catalyzed by GTP cyclohydrolase I.
Bracher
A
A
Lehrstuhl für Organische Chemie und Biochemie, Technische Universität München, Lichtenbergstrasse 4, D-85747 Garching, Germany. adelbert.bacher@ch.tum.de
Fischer
M
M
Eisenreich
W
W
Ritz
H
H
Schramek
N
N
Boyle
P
P
Gentili
P
P
Huber
R
R
Nar
H
H
Auerbach
G
G
Bacher
A
A
D020134:Catalytic Domain / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology; D006136:GTP Cyclohydrolase / Q000235:genetics / Q000378:metabolism*; D006160:Guanosine Triphosphate / Q000378:metabolism*; D006639:Histidine / Q000235:genetics; D008956:Models, Chemical; D009154:Mutation; D019798:Neopterin / Q000031:analogs & derivatives; D019906:Nuclear Magnetic Resonance, Biomolecular; D011621:Pteridines / Q000378:metabolism*; D011742:Pyrimidine Nucleotides / Q000378:metabolism*
C119702:2-amino-5-formylamino-6-ribofuranosylamino-4(3H)-pyrimidinone triphosphate; D011621:Pteridines; D011742:Pyrimidine Nucleotides; C013490:dihydroneopterin triphosphate; D006639:Histidine; D019798:Neopterin; D006160:Guanosine Triphosphate; D006136:GTP Cyclohydrolase