An endogeneous inhibitory factor for choline acetyltransferase

Abstract

Chromatography of partially purified choline acetyltransferase (CAT) over carboxymethyl cellulose may result in the loss of up to 95% of the enzyme activity. This loss of activity can be prevented by running the chromatographs at low protein concentration with a large gradient volume suggesting that interactions between CAT and other endogenous proteins are involved in the mechanism of inactivation. Further experiments showed that CM-cellulose chromatography separates an endogenous inhibitory factor(s) and an endogenous activating factor(s) which protects the enzyme from the action of the former. The inhibitory factor elutes with CAT and produces almost complete inactivation unless the protein concentration is maintained below 0.05 mg/ml. Mixing experiments demonstrated that the activating factor is capable of blocking the effect of the inhibitory factor. The low degree of temperature dependence of the inhibitory factor essentially rules out the possibility that the inhibitor is a proteolytic enzyme. The I50 was estimated to be 10(-7) M or less suggesting a possible physiological role of these factors in the regulation of CAT activity.