Chung C, Chung C D, Bieber L L
Department of Biochemistry, Michigan State University, East Lansing 48824.
Protein Expr Purif. 1991 Oct-Dec;2(5-6):426-31. doi: 10.1016/1046-5928(91)90104-q.
Heart and liver mitochondrial, as well as liver peroxisomal, carnitine acetyltransferase was purified to apparent homogeneity and some properties, primarily of heart mitochondrial carnitine acetyltransferase, were determined. Hill coefficients for propionyl-CoA are 1.0 for each of the enzymes. The molecular weight of heart mitochondrial carnitine acetyltransferase, determined by SDS-PAGE, is 62,000. It is monomeric in the presence of catalytic amounts of substrate. Polyclonal antibodies against purified rat liver peroxisomal carnitine acetyltransferase precipitate liver and heart mitochondrial and liver peroxisomal carnitine acetyltransferase, but not liver peroxisomal carnitine octanoyltransferase. Liver peroxisomes, mitochondria, and microsomes and heart mitochondria all give multiple bands on Western blotting with the antibody against carnitine acetyltransferase. Major protein bands occur at the molecular weight of carnitine acetyltransferase and at 33 to 35 kDa.
对心脏和肝脏线粒体以及肝脏过氧化物酶体中的肉碱乙酰转移酶进行了纯化，使其达到表观均一性，并测定了一些性质，主要是心脏线粒体肉碱乙酰转移酶的性质。每种酶对丙酰辅酶A的希尔系数均为1.0。通过SDS-PAGE测定，心脏线粒体肉碱乙酰转移酶的分子量为62,000。在存在催化量底物的情况下，它呈单体形式。针对纯化的大鼠肝脏过氧化物酶体肉碱乙酰转移酶制备的多克隆抗体可沉淀肝脏和心脏线粒体以及肝脏过氧化物酶体中的肉碱乙酰转移酶，但不能沉淀肝脏过氧化物酶体中的肉碱辛酰转移酶。肝脏过氧化物酶体、线粒体、微粒体以及心脏线粒体在用抗肉碱乙酰转移酶抗体进行蛋白质印迹分析时均出现多条条带。主要蛋白条带出现在肉碱乙酰转移酶的分子量处以及33至35 kDa处。
