Isolation of homologous and heterologous complexes between catalytic and regulatory components of adenylate cyclase by forskolin-Sepharose.
Homologous and heterologous complexes between catalytic and GTP-binding components can be isolated by means of immobilized succinyldeacetylforskolin (forskolin-Sepharose). A heterologous complex is formed by reconstitution of forskolin-Sepharose bound catalytic function from rabbit myocardial membranes with the homogenous [3H]methyl-GTP-binding protein from duck erythrocyte membranes. Analysis of the reconstituted complex by sodium dodecyl sulfate polyacrylamide gelelectrophoresis reveals that only the Mr 42 000 component of the GTP-binding protein's Mr 42 000/Mr 35 000 heterodimer contributes to the formation of active adenylate cyclase.
D000262:Adenylyl Cyclases / Q000302:isolation & purification* / Q000378:metabolism; D000818:Animals; D002846:Chromatography, Affinity; D005576:Colforsin; D004224:Diterpenes; D004372:Ducks; D004789:Enzyme Activation; D004910:Erythrocyte Membrane / Q000201:enzymology*; D019204:GTP-Binding Proteins; D007700:Kinetics; D009206:Myocardium / Q000201:enzymology*; D011817:Rabbits; D011956:Receptors, Cell Surface / Q000302:isolation & purification* / Q000378:metabolism; D012685:Sepharose
D004224:Diterpenes; D011956:Receptors, Cell Surface; D005576:Colforsin; D012685:Sepharose; D019204:GTP-Binding Proteins; D000262:Adenylyl Cyclases