Reversible activation of soluble guanylate cyclase by oxidizing agents.
Soluble guanylate cyclase of human platelets was stimulated by thiol oxidizing compounds like diamide and the reactive disulfide 4, 4'-dithiodipyridine. Activation followed a bell-shaped curve, revealing somewhat different optimum concentrations for each compound, although in both cases, higher concentrations were inhibitory. Diamide at a concentration of 100 microM transiently activated the enzyme. In the presence of moderate concentrations of diamide and 4,4'-dithiodipyridine, causing a two- to fourfold activation by themselves, the stimulatory activity of NO-releasing compounds like sodium nitroprusside was potentiated. In contrast, higher concentrations of thiol oxidizing compounds inhibited the NO-stimulated activation of soluble guanylate cyclase. Activation of guanylate cyclase was accompanied by a reduction in reduced glutathione and a concomitant formation of protein-bound glutathione (protein-SSG). Both compounds showed an activating potency as long as reduced glutathione remained, leading to inhibition of the enzyme just when all reduced glutathione was oxidized. Activation was reversible while reduced glutathione recovered and protein-SSG disappeared. We propose that diamide or reactive disulfides and other thiol oxidizing compounds inducing thiol-disulfide exchange activate soluble guanylate cyclase. In this respect partial oxidation is associated with enzyme activation, whereas massive oxidation results in loss of enzymatic activity. Physiologically, partial disulfide formation may amplify the signal toward NO as the endogenous activator of soluble guanylate cyclase.
Wu
X B
XB
University of Konstanz, Faculty of Biology, Federal Republic of Germany.
Brüne
B
B
von Appen
F
F
Ullrich
V
V
D001792:Blood Platelets / Q000201:enzymology*; D006152:Cyclic GMP / Q000097:blood; D003958:Diamide / Q000008:administration & dosage / Q000494:pharmacology; D004220:Disulfides / Q000008:administration & dosage / Q000097:blood / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D004789:Enzyme Activation / Q000187:drug effects; D005978:Glutathione / Q000097:blood; D006162:Guanylate Cyclase / Q000097:blood*; D006801:Humans; D007700:Kinetics; D009569:Nitric Oxide / Q000378:metabolism; D009599:Nitroprusside / Q000494:pharmacology; D016877:Oxidants / Q000494:pharmacology*; D010084:Oxidation-Reduction; D011725:Pyridines / Q000008:administration & dosage / Q000494:pharmacology; D013438:Sulfhydryl Compounds / Q000097:blood
D004220:Disulfides; D016877:Oxidants; D011725:Pyridines; D013438:Sulfhydryl Compounds; D003958:Diamide; D009599:Nitroprusside; C007542:4,4'-dipyridyl disulfide; D009569:Nitric Oxide; D006162:Guanylate Cyclase; D005978:Glutathione; D006152:Cyclic GMP