Solubilization of guanylyl cyclase from bovine rod outer segments and effects of lowering Ca2+ and nitro compounds.
Guanylyl cyclase from bovine rod outer segments was solubilized using Triton X-100 and a high concentration of KCl, and its regulation was studied. The efficiency of solubilization was about 50-90% of total activity. When the Ca2+ content was lowered (less than 80 nM), guanylyl cyclase was activated about 2-fold. In the presence of higher concentrations of Ca2+ (greater than 140 nM), the activity was decreased. The regulation by Ca2+ was also demonstrated with solubilized preparations. In the presence of 186 nM Ca2+ which inhibited guanylyl cyclase, La3+ activated the enzyme about 2-fold, suggesting that the Ca2(+)-binding protein similar to other Ca2(+)-binding proteins associates with guanylyl cyclase regulation. Sodium nitroprusside and nitric oxide which are activators of soluble guanylyl cyclase in other tissues also activated the retinal guanylyl cyclase. Maximum activation by sodium nitroprusside was 20-fold using Mg2+ as a cofactor. Activation by nitric oxide and related compounds suggests that retinal guanylyl cyclase contains a heme prosthetic group that may participate in a novel regulatory mechanism for this enzyme.
Horio
Y
Y
Department of Pharmacology, Northwestern University Medical School, Chicago, Illinois 60611.
Murad
F
F
D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D002413:Cations, Divalent; D002417:Cattle; D003902:Detergents; D004789:Enzyme Activation; D006162:Guanylate Cyclase / Q000737:chemistry* / Q000378:metabolism; D009569:Nitric Oxide / Q000494:pharmacology; D009574:Nitro Compounds / Q000494:pharmacology*; D009599:Nitroprusside / Q000494:pharmacology; D017830:Octoxynol; D011092:Polyethylene Glycols; D012374:Rod Cell Outer Segment / Q000187:drug effects / Q000201:enzymology*
D002413:Cations, Divalent; D003902:Detergents; D009574:Nitro Compounds; D009599:Nitroprusside; D009569:Nitric Oxide; D011092:Polyethylene Glycols; C005092:lubrol; D017830:Octoxynol; D006162:Guanylate Cyclase; D002118:Calcium