Role of conformational changes in the heme-dependent regulation of human soluble guanylate cyclase.
Soluble guanylate cyclase (sGC) is a receptor for endogenous and exogenous nitric oxide (NO) and is activated many fold upon its binding, making it a core enzyme in the nitric oxide signal transduction pathway. Much effort has been made to understand the link between binding of NO at the sGC heme and activation of the cyclase activity. We report here the first direct evidence for the role of conformational changes in transmitting the signal between the heme and cyclase domains. Using both circular dichroism (CD) and fluorescence spectroscopies, we have probed the effect that the sGC activators NO and 3-(5'-hydroxymethyl-2'-furyl)-1-benzyl-indazole (YC-1) and the inhibitor 1H-[1,2,4]-oxadiazolo-[4,3-a]-quinoxalin-1-one (ODQ) have on the structure of the protein. Surprisingly, binding of either ODQ or YC-1 to NO-bound sGC cause virtually identical changes in the far-UV CD spectra of sGC, reflecting a perturbation in the secondary structure of the enzyme. This change is absent upon binding of NO, YC-1 or ODQ alone. Using this and previous data, we propose a working model for the mechanism of activation of sGC by NO and YC-1 and inhibition by ODQ.
Kosarikov
D N
DN
Department of Chemistry and Biochemistry, San Francisco State University, 1600 Holloway Avenue, San Francisco, CA 94132-4163, USA.
Lee
J M
JM
Uversky
V N
VN
Counts Gerber
N
N
D020106:Acrylamide; D002942:Circular Dichroism; D004789:Enzyme Activation; D006162:Guanylate Cyclase / Q000037:antagonists & inhibitors / Q000737:chemistry*; D006418:Heme / Q000737:chemistry*; D006801:Humans; D007191:Indazoles / Q000737:chemistry; D008956:Models, Chemical; D008958:Models, Molecular; D009569:Nitric Oxide / Q000737:chemistry; D020030:Nitric Oxide Donors / Q000737:chemistry; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D013050:Spectrometry, Fluorescence; D014364:Tryptophan
D007191:Indazoles; D020030:Nitric Oxide Donors; C090937:3-(5'-hydroxymethyl-2'-furyl)-1-benzylindazole; D020106:Acrylamide; D009569:Nitric Oxide; D006418:Heme; D014364:Tryptophan; D006162:Guanylate Cyclase