title: Effects of thiols, sugars, and proteins on nitric oxide activation of guanylate cyclase.
abstract: Purification of soluble guanylate cyclase from rat liver resulted in an apparent loss of enzyme activation by nitric oxide that could be restored by dithiothreitol. methemoglobin, bovine serum albumin, or sucrose. Although hemoglobin also permitted some activation with nitric oxide, the effect of other agents to restore enzyme activation was prevented with hemoglobin. As a result of enzyme purification, there is an alteration of the dose-response relationship for nitric oxide activation. After partial enzyme purification, relatively high concentrations of nitric oxide that were stimulatory in crude enzyme preparations had no effect on enzyme activity. However, partially purified or homogeneous enzyme was activated by lower concentrations of nitric oxide. The bell-shaped dose-response curve for nitric oxide was shifted to the left with guanylate cyclase purification. The addition of dithiothreitol, methemoglobin, bovine serum albumin, or sucrose to enzyme markedly broadens the dose-response curve for nitric oxide. Thus, the apparent loss of responsiveness to nitric oxide with purification is a function of increased sensitivity of guanylate cyclase to nitric oxide. Increased sensitivity to nitric oxide with enzyme purification probably results from the removal of heme, proteins, and small molecules that can serve as scavengers or sinks for nitric oxide and prevent excessive oxidation of the enzyme.
mesh_terms: D000818:Animals; D004229:Dithiothreitol / Q000494:pharmacology*; D004789:Enzyme Activation / Q000187:drug effects; D006162:Guanylate Cyclase / Q000378:metabolism*; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008706:Methemoglobin / Q000494:pharmacology*; D009569:Nitric Oxide / Q000494:pharmacology*; D051381:Rats; D012710:Serum Albumin, Bovine / Q000494:pharmacology*; D013395:Sucrose / Q000494:pharmacology*
chemical_list: D012710:Serum Albumin, Bovine; D009569:Nitric Oxide; D013395:Sucrose; D008706:Methemoglobin; D006162:Guanylate Cyclase; D004229:Dithiothreitol