Synergistic activation of soluble guanylate cyclase by YC-1 and carbon monoxide: implications for the role of cleavage of the iron-histidine bond during activation by nitric oxide.
BACKGROUND
Nitric oxide (.NO) is used in biology as both an intercellular signaling agent and a cytotoxic agent. In signaling, submicromolar quantities of .NO stimulate the soluble isoform of guanylate cyclase (sGC) in the receptor cell. .NO increases the Vmax of this heterodimeric hemoprotein up to 400-fold by interacting with the heme moiety of sGC to form a 5-coordinate complex. Carbon monoxide (CO) binds to the heme to form a 6-coordinate complex, but only activates the enzyme 5-fold, YC-1 is a recently discovered compound that relaxes vascular smooth muscle by stimulating sGC.


RESULTS
In the presence of YC-1, CO activates sGC to the same specific activity as attained with .NO. YC-1 did not affect the NO-stimulated activity. The on-rate (kon) and off-rate (koff) of CO for binding to sGC in the presence of YC-1 were determined by stopped-flow spectrophotometry. Neither the kon nor the koff varied from values previously obtained in the absence of YC-1, indicating that YC-1 has no effect on the affinity of CO for the heme. In the presence of YC-1, the visible spectrum of the sGC-CO complex has a Soret peak at 423 nm, indicating the complex is 6-coordinate.


CONCLUSIONS
YC-1 has no effect on the affinity of CO for the heme of sGC. In the presence of YC-1, maximal activation of sGC by CO is achieved by formation of a 6-coordinate complex between CO and the heme indicating that cleavage of the Fe-His bond is not required for maximal activation of sGC.
Stone
J R
JR
Department of Biological Chemistry, School of Medicine, University of Michigan, Ann Arbor 48109-1065, USA.
Marletta
M A
MA
D002248:Carbon Monoxide / Q000494:pharmacology*; D004789:Enzyme Activation; D006162:Guanylate Cyclase / Q000378:metabolism*; D006639:Histidine / Q000378:metabolism*; D006868:Hydrolysis; D007191:Indazoles / Q000494:pharmacology*; D007501:Iron / Q000378:metabolism*; D007700:Kinetics; D009569:Nitric Oxide / Q000494:pharmacology*
D007191:Indazoles; C090937:3-(5'-hydroxymethyl-2'-furyl)-1-benzylindazole; D009569:Nitric Oxide; D006639:Histidine; D002248:Carbon Monoxide; D007501:Iron; D006162:Guanylate Cyclase