A sensitive method for the assay of guanylate cyclase activity.
A method for the assay of guanylate cyclase is described utilizing alpha-[32P]-GTP as substrate for the enzyme reaction. 100-150 microgram of enzyme protein is incubated in a 15.6 mM Tris-HCl buffer incubation mixture, pH 7.6. The reaction is stopped by the addition of EDTA. The [32P]-cyclic GMP formed is separated by a two-step column chromatography on Dowex 50W-X4 ion-exchange resin and neutral alumina. The recovery for cyclic GMP was about 70%. The blank values ranged from 0.001-0.003% of the added alpha-[32P]-GTP which had been purified by Dowex 50W-X4 column chromatography. This method was employed for the assay of guanylate cyclase activities in different tissues.
D000818:Animals; D001792:Blood Platelets / Q000201:enzymology; D002852:Chromatography, Ion Exchange / Q000379:methods; D006162:Guanylate Cyclase / Q000032:analysis*; D006168:Guinea Pigs; D006801:Humans; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008168:Lung / Q000201:enzymology; D008297:Male; D008832:Microchemistry; D009206:Myocardium / Q000201:enzymology; D009928:Organ Specificity; D051381:Rats; D013045:Species Specificity
D006162:Guanylate Cyclase