Pterin interactions with distinct reductase activities of NO synthase.
Besides oxidizing L-arginine, neuronal NO synthase (NOS) NADPH-dependently reduces various electron acceptors, including cytochrome c and tetrazolium salts. The latter NADPH diaphorase reaction is used as a NOS-specific histochemical stain. Both reductase activities have been utilized to analyse electron transfer mechanisms within NOS. Basal L-arginine turnover by homodimeric NOS is enhanced by exogenous tetrahydrobiopterin, and the intra-subunit electron flow may include intermediate trihydrobiopterin. In the present work we have investigated the possible role of the tetrahydrobiopterin binding site of NOS in its reductase activities by examining the effects of anti-pterin type (PHS) NOS inhibitors. Although the type I anti-pterin, PHS-32, which does not affect basal dimeric NOS activity, also had no effect on either reductase activity, the type II anti-pterin, PHS-72, which inhibits basal NOS activity, inhibited both reductase activities and the NADPH diaphorase histochemical stain. Pterin-free NOS monomers catalysed both cytochrome c and tetrazolium salt reduction. Our data suggest that both NOS reductase activities are independent of tetrahydrobiopterin. However, occupation of an exosite near the pterin site in NOS by type II anti-pterins may interfere with the electron flow within the active centre, suggesting that steric perturbation of the pterin binding pocket or reductase interaction contribute to the mechanism of inhibition by this class of NOS inhibitors.
Pantke
M M
MM
Department of Pharmacology and Toxicology, Julius-Maximilians-University Würzburg, Versbacher Str. 9, D-97078 Würzburg, Germany.
Reif
A
A
Valtschanoff
J G
JG
Shutenko
Z
Z
Frey
A
A
Weinberg
R J
RJ
Pfleiderer
W
W
Schmidt
H H
HH
D001708:Biopterins / Q000031:analogs & derivatives* / Q000494:pharmacology*; D002118:Calcium / Q000494:pharmacology; D002147:Calmodulin / Q000494:pharmacology; D002374:Catalase / Q000378:metabolism; D004579:Electron Transport; D009245:NADH Dehydrogenase / Q000378:metabolism; D009252:NADPH Dehydrogenase / Q000187:drug effects*; D009569:Nitric Oxide / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000187:drug effects*; D052247:Nitric Oxide Synthase Type II; D010084:Oxidation-Reduction; D010088:Oxidoreductases; D011622:Pterins / Q000037:antagonists & inhibitors; D013482:Superoxide Dismutase / Q000378:metabolism; D013481:Superoxides / Q000378:metabolism; D013778:Tetrazolium Salts / Q000378:metabolism
D002147:Calmodulin; D011622:Pterins; D013778:Tetrazolium Salts; D013481:Superoxides; D001708:Biopterins; D009569:Nitric Oxide; D010088:Oxidoreductases; D002374:Catalase; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; D013482:Superoxide Dismutase; D009252:NADPH Dehydrogenase; D009245:NADH Dehydrogenase; C034747:nitric-oxide reductase; C003402:sapropterin; D002118:Calcium