Cholinesterase activity of the motor endplate in isolated muscle membrane.

Abstract

Abstract— The cholinesterase activity of motor endplates in tibialis anterior muscle of rats accounted for about 20 per cent of the total cholinesterase activity of the muscle. In the isolated muscle membrane preparation of rat intercostal muscle, the cholinesterase activity was localized solely in the motor endplate, as shown by cholinesterase staining. The cholinesterase activity of the membrane per unit of nitrogen was 26·9 times that of the muscle homogenate. The membrane (endplate) cholinesterase had an optimal pH of 8, Km value of 3·1 mm, and was stable at 4° for at least 13 days. Cholinesterase of a motor endplate hydrolysed 2·69 × 108 acetylcholine molecules in 1 msec. Since it is estimated that 108 cholinesterase active sites are present in a motor endplate, the turnover time (time necessary for one enzyme site to hydrolyse one acetylcholine molecule) is calculated to be 372 μsec, and the turnover number (molecules of acetylcholine hydrolysed by one enzyme site/min) to be 1·61 × 105. From studies with cholinesterase inhibitors, cholinesterase activity was estimated to be due mostly to acetylcholinesterase, and only a minor part to pseudocholinesterase. The muscle membrane preparation seems to be useful for the study of other properties of the motor endplate.