PURIFICATION AND CHARACTERIZATION OF 2-SUBUNIT CYTOCHROME AA3 FROM BACILLUS-CEREUS

被引：11

作者：

GARCIAHORSMAN, JA ^{[1
]}

BARQUERA, B ^{[1
]}

GONZALEZHALPHEN, D ^{[1
]}

ESCAMILLA, JE ^{[1
]}

机构：

[1] NATL AUTONOMOUS UNIV MEXICO,INST FISIOL CELULAR,DEPT MICROBIOL,APARTADO POSTAL 70-242,MEXICO CITY 04510,DF,MEXICO

来源：

MOLECULAR MICROBIOLOGY | 1991年 / 5卷 / 01期

关键词：

D O I：

10.1111/j.1365-2958.1991.tb01840.x

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Cytochrome c-oxidase type aa3 (EC 1.9.3.1) was purified to homogeneity from vegetative Bacillus cereus by ion-exchange and hydroxylapatite chromatography in the presence of Triton X-100. Gel filtration analysis suggested a dimeric structure apparently 172kDa in size; however, only a monomer of 81kDa was detected when analysed by non-denaturing gel electrophoresis. Denaturing gel electrophoresis analysis of the protein showed the presence of two subunits (51 and 30kDa). Atomic adsorption and visible spectroscopy showed typical aa3 redox centres with haem a iron and copper in a ratio of 22nmol and 35ng-atom per mg protein, respectively. No haem c was found associated with the purified enzyme in the conditions reported here. Oxidase activity was fully reconstituted by phospholipids in the presence of N,N,N',N'-tetramethyl-p-phenylenediamine or reduced yeast cytochrome c (but not horse cytochrome c) as electron donors. This activity was abolished by cyanide and carbon monoxide.

引用

页码：197 / 205

页数：9

共 40 条

[1] COMPLETE SEQUENCE OF BOVINE MITOCHONDRIAL-DNA - CONSERVED FEATURES OF THE MAMMALIAN MITOCHONDRIAL GENOME [J].