Subject(s)
Anions/pharmacology , Guanylate Cyclase/antagonists & inhibitors , Lung/enzymology , Sulfates/pharmacology , Animals , Carrageenan/pharmacology , Dextran Sulfate , Dextrans/pharmacology , Dose-Response Relationship, Drug , Heparin/pharmacology , Manganese/pharmacology , Polyanetholesulfonate/pharmacology , Polyvinyls/pharmacology , RatsABSTRACT
Soluble guanylate cyclase (GTP pyrophosphate-lyase (cyclizing), EC 4.6.1.2) from rat lung demonstrated concentration-dependent stimulation, that is, an increase in specific activity with increasing enzyme (protein) concentration. This phenomenon persisted through several steps of enzyme purification and was apparently due to the presence of a macromolecular activator, similar in size to the enzyme. Treatment of partially purified enzyme with N-ethylmaleimide destroyed catalytic activity, but did not effect the ability of the preparation to stimulate activity. Kinetic analysis demonstrated that the stimulation was due to an increased V value with no change in the apparent Km value for MnGTP. Stimulation occurred without a time lag, the activator apparently interacting reversibly with the enzyme to increase catalytic capability. Some nonionic detergents of the Triton series inhibited enzyme activity by decreasing the V value, with no change in the Km value, and also decreased concentration-dependent stimulation. However, the two phenomena were not directly related. While the physiological significance of the activator is unclear, its presence affects estimations of recovery during enzyme purification, V determinations, and determinations of the effect of hormone or drug treatment on the activity of tissue extracts.
