McIntyre J O, Latruffe N, Brenner S C, Fleischer S
Department of Molecular Biology, Vanderbilt University, Nashville, Tennessee 37235.
Arch Biochem Biophys. 1988 Apr;262(1):85-98. doi: 10.1016/0003-9861(88)90171-3.
3-Hydroxybutyrate dehydrogenase is a lipid-requiring enzyme with an absolute requirement of phosphatidylcholine for enzymatic activity. Purification of the enzyme to homogeneity from bovine heart mitochondria was described more than a decade ago [H. G. Bock and S. Fleischer (1975) J. Biol. Chem. 250, 5774-5781]. We have modified the purification procedure so that it is faster, the yield has been improved, and the specific activity is greater by approximately 50%. The updated procedure has also been applied to isolate the enzyme from rat liver mitochondria. Characteristics of the enzyme from bovine heart and rat liver mitochondria have been compared and found to be similar with respect to: (1) purification characteristics; (2) amino acid composition; (3) pH optimum for enzymatic activity; (4) kinetic characteristics; (5) molecular weight as determined by sedimentation equilibrium in guanidine hydrochloride; (6) peptide maps; (7) immunological cross-reactivity. These studies show that 3-hydroxybutyrate dehydrogenase from bovine heart and rat liver mitochondria, though similar, are not identical.
3-羟基丁酸脱氢酶是一种需要脂质的酶，其酶活性绝对需要磷脂酰胆碱。十多年前就描述了从牛心线粒体中将该酶纯化至同质的方法[H.G.博克和S.弗莱舍尔（1975年）《生物化学杂志》250, 5774 - 5781]。我们改进了纯化程序，使其更快，产量有所提高，比活性提高了约50%。更新后的程序也已应用于从大鼠肝线粒体中分离该酶。已对牛心和大鼠肝线粒体中的酶的特性进行了比较，发现它们在以下方面相似：（1）纯化特性；（2）氨基酸组成；（3）酶活性的最适pH值；（4）动力学特性；（5）通过在盐酸胍中沉降平衡测定的分子量；（6）肽图；（7）免疫交叉反应性。这些研究表明，牛心和大鼠肝线粒体中的3-羟基丁酸脱氢酶虽然相似，但并不相同。
