细胞色素C氧化酶的纯化及动力学性质研究  

Purification and kinetics of Cytochrome C Oxidase

作　　者：李丹彤[1] 刘愉[2] 邬玉净[1] 冯雁[3] 杨同书[3] 

机构地区：[1]大连水产学院养殖系,辽宁大连116023 [2]长春市儿童医院,吉林长春130021 [3]吉林大学酶工程重点实验室,吉林长春130023

出　　处：《大连水产学院学报》2001年第2期112-117,共6页Journal of Dalian Fisheries University

摘　　要：采用硫酸铵分级沉淀和CytochromeC -Sepharose 4B亲和层析 ,从牛心线粒体中纯化了细胞色素C氧化酶 (CytochromeCOxidase ,简称CCO ) ,其比活性为 1 7 88s- 1 ·mg- 1 ,聚丙烯酰胺凝胶电泳为一条带 ,血红素A (HemeaA)含量达到 1 5 4nmol/mg。纯化后的CCO具有典型的CCO特征 ,还原态CCO峰值出现在 60 5nm和 443nm ,氧化态CCO峰值出现在 598nm和 42 4nm。CCO的稳态动力学参数Vm1、Km1、Vm2 、Km2 分别为 1 42s- 1 、 0 53μmol/L、 2 71s- 1 、 1 81 μmol/L。The Cytochrome C Oxidase (CCO) had been purified by ammonium sulfate fractionation and an affinity chromatography on the cytochrome C-Sepharose 4B column. Purified CCO showed one single band on PAGE. Specific activity was 17 88 s -1 ·mg -1 . Heme A content was 15 4 nmol/mg. It showed typical characteristics of CCO. Peak absorption of reduced form was at 605 nm and 443 nm. Peak absorption of oxidative form was at 598 nm and 424 nm. The CCO kinetic parameters \%V\%\-\{m 1\}, \%K\%\-\{m 1\}, \%V\%\-\{m 2\} and \%K\%\-\{m 2\} were 1 42 s -1 , 0 53 μmol/L, 2 71 s -1 and 1 81 μmol/L, respectively.

关 键 词：细胞色素C氧化酶 纯化 动力学 参数 线粒体病 

分 类 号：Q554.9[生物学—生物化学]