Murthy M S, Kamanna V S, Pande S V
Biochem J. 1986 May 15;236(1):143-8. doi: 10.1042/bj2360143.
A simple method for assaying the mitochondrial carnitine/acylcarnitine translocase of muscles that needs only few milligrams of fresh tissue is described. The procedure involves monitoring of the sulphobetaine (an inhibitor of the translocase)-sensitive acetylation of sub-saturating concentrations of carnitine in the medium, linked to the oxidation of [2-14C]pyruvate in the presence of malonate. Conditions affecting the reliability of the outlined procedure and the ancillary information to be collected, namely the activities of pyruvate oxidase system and carnitine acetyltransferase, for detecting possible deficiency of the translocase are described, together with data on the translocase activity in human skeletal muscle, in rat red and white skeletal muscles and in rat heart. The concepts outlined should allow development of assays of other mitochondrial transporters that also would require neither isolation of mitochondria nor availability of a large quantity of tissue, both of which are otherwise needed at present.
本文描述了一种仅需几毫克新鲜组织就能测定肌肉线粒体肉碱/酰基肉碱转位酶的简单方法。该方法包括监测磺基甜菜碱（转位酶抑制剂）对培养基中亚饱和浓度肉碱乙酰化的敏感性，此过程与丙二酸存在下[2-¹⁴C]丙酮酸的氧化相关。文中描述了影响所概述方法可靠性的条件以及为检测转位酶可能缺乏而需收集的辅助信息，即丙酮酸氧化酶系统和肉碱乙酰转移酶的活性，同时还给出了人骨骼肌、大鼠红白骨骼肌及大鼠心脏中转位酶活性的数据。所概述的概念应有助于开发其他线粒体转运体的检测方法，这些方法目前既不需要分离线粒体，也不需要大量组织。
