title: Kinetics and equilibria of soluble guanylate cyclase ligation by CO: effect of YC-1.
abstract: Previous work has proved that the enzyme-soluble guanylate cyclase, GC, is activated several 100-fold by the combination of carbon monoxide plus a benzylindazole derivative called YC-1. That is about the same as activation by nitric oxide, which has a well-established role both in vivo and in vitro. This report addresses several spectroscopic, equilibrium, and kinetic effects wrought by YC-1 on carboxyl guanylate cyclase, including the following: a shift in the Soret absorption band by 4 nm to shorter wavelength; an increase in CO affinity by an order of magnitude; a dramatic change in the kinetics of CO association. After photolytic dissociation of CO, the majority, but not all, of bimolecular ligand recombination occurs with a time constant about 1000-fold faster than in the absence of YC-1, while a smaller fraction recombines almost, but not quite, the same as usual. This is reminiscent of the kinetics of NO association with GC, which also shows two prominent phases. The results just listed pertain in the presence of GTP/cGMP, which would be present during enzyme catalysis. Qualitatively similar, but smaller, effects occur in the absence of GTP/cGMP. Measurements are reported to characterize other changes in buffer conditions. The results are consistent with a mechanistic model that attributes a crucial role to the proximal bond that connects the heme iron to a histidine side chain in GC but also requires protein control of the distal environment.
authors.affiliation: Department of Chemistry, University of California-San Diego, La Jolla 92093, USA.
authors.forename: V G
authors.initials: VG
authors.lastname: Kharitonov
authors.forename: V S
authors.initials: VS
authors.lastname: Sharma
authors.forename: D
authors.initials: D
authors.lastname: Magde
authors.forename: D
authors.initials: D
authors.lastname: Koesling
mesh_terms: D000818:Animals; D002248:Carbon Monoxide / Q000737:chemistry* / Q000378:metabolism*; D002417:Cattle; D006160:Guanosine Triphosphate / Q000737:chemistry / Q000378:metabolism; D006162:Guanylate Cyclase / Q000737:chemistry* / Q000378:metabolism*; D007191:Indazoles / Q000737:chemistry* / Q000378:metabolism*; D007700:Kinetics; D008024:Ligands; D008956:Models, Chemical; D009569:Nitric Oxide / Q000737:chemistry / Q000378:metabolism; D010782:Photolysis; D010975:Platelet Aggregation Inhibitors / Q000737:chemistry / Q000378:metabolism; D012995:Solubility; D013056:Spectrophotometry, Ultraviolet
chemical_list: D007191:Indazoles; D008024:Ligands; D010975:Platelet Aggregation Inhibitors; C090937:3-(5'-hydroxymethyl-2'-furyl)-1-benzylindazole; D009569:Nitric Oxide; D002248:Carbon Monoxide; D006160:Guanosine Triphosphate; D006162:Guanylate Cyclase