Characterization of adenylate cyclase purified from rat brain by hydrophobic chromatography

Hydrophobic chromatography of detergent-solubilized rat brain adenylate cyclase on dodecyl-Sepharose produced a species that was soluble in the absence of detergent and could be manipulated like a conventional hydrophilic protein. Sevenfold purification was achieved by this technique. Further purification could then be effected by affinity chromatography on ATP-Sepharose. The purified enzyme was no longer sensitive to fluoride or guanyl nucleotides. No interaction of brain adenylate cyclase was observed with immobilized triazinyl dyes such as Cibacron Blue 3GA nor with con-canavalin A-Sepharose. The molecular weight of the fluoride-activated catalytic complex in a freeze-dried membrane preparation was estimated to be 133,000 by irradiation inactivation.