The role of tetrahydrobiopterin in the activation of oxygen by nitric-oxide synthase.
We have studied the reaction of reduced nitric-oxide synthase (NOS) with molecular oxygen at -30 degrees C. In the first reaction cycle (from L-Arg to hydroxy-L-Arg), an oxygen adduct complex formed rapidly. Experiments in the absence of the reductase domain demonstrated that this complex was then further reduced by one electron stemming from the cofactor tetrahydrobiopterin (BH4). Spectral evidence suggested an iron(IV) porphyrin pi-cation radical as an intermediate. The nature of the oxidized BH4 was identified by EPR as a BH3* radical. Within the second cycle (from hydroxy-L-Arg to citrulline and NO), an iron(III)-NO complex could be identified clearly by its spectral characteristics. The strict requirement of BH4 for its formation suggests that BH4 plays a redox role, although transient, also in the second reaction cycle.
Bec
N
N
lnstitut für Pharmakologie und Toxikologie, Karl-Franzens-Universität, Graz, Austria.
Gorren AFC
Mayer
B
B
Schmidt
P P
PP
Andersson
K K
KK
Lange
R
R
D001708:Biopterins / Q000031:analogs & derivatives* / Q000737:chemistry* / Q000502:physiology*; D004578:Electron Spin Resonance Spectroscopy; D004583:Electrons; D008956:Models, Chemical; D019001:Nitric Oxide Synthase / Q000737:chemistry* / Q000378:metabolism; D052248:Nitric Oxide Synthase Type I; D052250:Nitric Oxide Synthase Type III; D010084:Oxidation-Reduction; D010100:Oxygen / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D013696:Temperature
D001708:Biopterins; D019001:Nitric Oxide Synthase; D052248:Nitric Oxide Synthase Type I; D052250:Nitric Oxide Synthase Type III; C003402:sapropterin; D010100:Oxygen