Immunological characterization of the cytochrome o terminal oxidase from Escherichia coli.
The cytochrome o terminal oxidase from Escherichia coli was immunochemically purified and monospecific antiserum toward cytochrome o was obtained. This antiserum is able to precipitate 100% of the ubiquinol-1 oxidase activity in Triton X-100 extracts of membranes from an E. coli strain in which cytochrome o is the only terminal oxidase. Cytochrome o was analyzed and quantitated using crossed immunoelectrophoresis, rocket immunoelectrophoresis, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis shows that cytochrome o is composed of four subunits of approximate equimolar stoichiometry with molecular weights of 51,000, 28,500, 18,000, and 12,700. The low temperature (77 K) reduced - oxidized spectrum of the immunoprecipitate shows two peaks at 555 and 562 nm, indicating b-type cytochromes. With the anti-cytochrome o and antiserum toward the cytochrome d terminal oxidase complex which was previously obtained, it is possible to immunochemically assay for all the cytochromes in the cytoplasmic membrane of aerobically grown E. coli. Preliminary results indicate that the biosynthesis of cytochrome o is repressed when cytochrome d is induced by lowering the dissolved oxygen concentration during cell growth.
D003573:Cytochrome b Group / Q000032:analysis; D003580:Cytochromes / Q000276:immunology*; D004926:Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins; D007122:Immunoelectrophoresis; D046911:Macromolecular Substances; D008970:Molecular Weight; D017830:Octoxynol; D011092:Polyethylene Glycols
D003573:Cytochrome b Group; D003580:Cytochromes; D029968:Escherichia coli Proteins; D046911:Macromolecular Substances; D011092:Polyethylene Glycols; D017830:Octoxynol; C017168:cytochrome bo, E coli; C017850:cytochrome b562, E coli