Catecholamine-sensitive guanylate cyclase from human caudate nucleus.
Partial purification of soluble guanylate cyclase on DEAE-Sephacel yields two separate peaks of guanylate cyclase activity. After 10-fold purification of the soluble enzyme, guanylate cyclase is markedly inhibited by micromolar concentrations of dopamine (I50 = 0.2 microM). Dopamine inhibition is observed whether the reaction is conducted with Mn2+ or with Mg2+, under atmosphere or N2(g), and using enzyme from either peak from the DEAE-Sephacel column. Other catecholamines also inhibit partially purified guanylate cyclase with an order of potency at 1 microM of: dopamine = L-DOPA > norepinephrine = isoproterenol = adrenochrome > epinephrine. The structural requirements for inhibition are two free hydroxyl groups on the phenyl ring and an ethylamine side chain. Dopamine also inhibits the Triton X-100-solubilized microsomal guanylate cyclase after partial purification on DEAE-Sephacel. Neither chlorpromazine, propranolol, nor phentolamine at 20 microM effectively block the dopamine inhibition of partially purified soluble guanylate cyclase. Micromolar concentrations of the reducing agents dithiothreitol and glutathione also inhibit partially purified guanylate cyclase, but unlike these agents, catecholamines can inhibit whether added in the reduced or the oxidized forms. Inhibition of enzyme activity by micromolar concentrations of dopamine, adrenochrome, or dithiothreitol is rapidly reversed by dilution and the dopamine inhibition is competitive with MgGTP. Inhibition does not appear to involve covalent binding or to result from the ability of catecholamines to reduce the concentrations of oxygen or free radicals in solution.
D000323:Adrenochrome / Q000494:pharmacology; D002395:Catecholamines / Q000494:pharmacology*; D002421:Caudate Nucleus / Q000201:enzymology*; D004229:Dithiothreitol / Q000494:pharmacology; D004298:Dopamine / Q000494:pharmacology; D006162:Guanylate Cyclase / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D013329:Structure-Activity Relationship
D002395:Catecholamines; D000323:Adrenochrome; D006162:Guanylate Cyclase; D004229:Dithiothreitol; D004298:Dopamine