Activation of rat liver guanylate cyclase by proteolysis.
Guanylate cyclase activity (GTP pyrophosphate-lyase (cyclizing), EC 4.6.1.2.), measured in purified rat liver plasma membranes, was markedly increased by treatment with various purified proteases. The effect was maximal with trypsin, alpha-chymotrypsin, papain, and thermolysin (6- to 8-fold increase with 5 to 20 microgram of protease/ml) and lower with subtilisin and elastase (3- to 4-fold increase). The activation was due to an increase in the maximal velocity of the cyclizing reaction. No modification was observed either in the apparent affinity for the substrate MnGTP or in the cooperative behavior of the enzyme kinetics which displayed Hill coefficients of 1.6 for both basal and activated states. The Triton X-100-dispersed guanylate cyclase remained sensitive to papain, which suggests that the action of proteases was not restricted to an indirect action upon the membranous environment of the guanylate cyclase. In contrast, the cytosolic soluble guanylate cyclase, assayed in the presence or absence of sodium azide, was absolutely insensitive to papain. Thus, proteolysis represents a previously undescribed mechanism for activating membranous guanylate cyclase systems, which might be of importance in the physiological regulation of this enzyme.
D000818:Animals; D002918:Chymotrypsin / Q000494:pharmacology; D010450:Endopeptidases / Q000494:pharmacology*; D004789:Enzyme Activation; D005260:Female; D006160:Guanosine Triphosphate / Q000494:pharmacology; D006162:Guanylate Cyclase / Q000378:metabolism*; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008345:Manganese / Q000494:pharmacology; D010196:Pancreatic Elastase / Q000494:pharmacology; D010206:Papain / Q000494:pharmacology; D051381:Rats; D013381:Subtilisins / Q000494:pharmacology; D013820:Thermolysin / Q000494:pharmacology; D014357:Trypsin / Q000494:pharmacology
D008345:Manganese; D006160:Guanosine Triphosphate; D010450:Endopeptidases; D013381:Subtilisins; D002918:Chymotrypsin; D010196:Pancreatic Elastase; D014357:Trypsin; D010206:Papain; D013820:Thermolysin; D006162:Guanylate Cyclase