title: Variable forms of soluble guanylyl cyclase: protein-ligand interactions and the issue of activation by carbon monoxide.
abstract: Soluble guanylyl cyclase (sGC) is known to be activated by NO binding to the heme moiety; previous studies have shown that CO does not activate sGC to the same extent as NO. Resonance Raman spectroscopy reveals different heme pocket structures for soluble guanylyl cyclase prepared by alternate methods, all of which display activation by NO. In our preparation, and in the expressed protein sGC1, the resting Fe(II) state is mainly 6-coordinate and low-spin, and the CO adduct has vibrational frequencies characteristic of a histidine-heme-CO complex in a hydrophobic environment. In contrast, the protein sGC2 is 5-coordinate, high-spin in the resting state, and the CO adduct has perturbed vibrational frequencies indicative of a negatively polarizing residue in the binding pocket. The differences may result from the need to reconstitute sGC1 or different isolation procedures for sGC1 versus sGC2. However, both sGC1 and sGC2 are activated by the same mechanism, namely displacement of the proximal histidine ligand upon NO binding, and neither one is activated by CO. If CO is an activator in vivo, some additional molecular component is required.
authors.affiliation: Department of Chemistry, Princeton University, NJ 08544, USA.
authors.forename: K M
authors.initials: KM
authors.lastname: Vogel
authors.forename: S
authors.initials: S
authors.lastname: Hu
authors.forename: T G
authors.initials: TG
authors.lastname: Spiro
authors.forename: E A
authors.initials: EA
authors.lastname: Dierks
authors.forename: A E
authors.initials: AE
authors.lastname: Yu
authors.forename: J N
authors.initials: JN
authors.lastname: Burstyn
mesh_terms: D000818:Animals; D002248:Carbon Monoxide / Q000378:metabolism*; D002417:Cattle; D004789:Enzyme Activation; D006162:Guanylate Cyclase / Q000737:chemistry / Q000378:metabolism*; D007527:Isoenzymes / Q000737:chemistry / Q000378:metabolism*; D008168:Lung / Q000201:enzymology; D009569:Nitric Oxide / Q000378:metabolism; D012995:Solubility; D013059:Spectrum Analysis, Raman
chemical_list: D007527:Isoenzymes; D009569:Nitric Oxide; D002248:Carbon Monoxide; D006162:Guanylate Cyclase