Analysis of the topology of the cytochrome d terminal oxidase complex of Escherichia coli by alkaline phosphatase fusions.
The cytochrome d complex of Escherichia coli is a heterodimer located in the bacterial cytoplasmic membrane, where it functions as a terminal oxidase of the aerobic respiratory chain. The topology of each of the two subunits of the cytochrome d complex was analysed by the genetic method involving alkaline phosphatase gene fusions. These fusions were generated by both an in vivo method using the transposon TnphoA and an in vitro method of construction. A total of 48 unique fusions were isolated and the whole-cell alkaline phosphatase-specific activities were determined. Data from these fusions, in combination with information from other studies, provide the basis for two-dimensional models for each of the two subunits, defining the way in which the subunits fold in the inner membrane of E. coli.
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School of Chemical Sciences, University of Illinois, Urbana 61801.
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Gennis
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D000469:Alkaline Phosphatase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D003573:Cytochrome b Group; D017937:Cytochrome d Group; D003580:Cytochromes / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D004251:DNA Transposable Elements; D045222:Electron Transport Chain Complex Proteins; D000939:Epitopes / Q000032:analysis; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D029968:Escherichia coli Proteins; D005798:Genes, Bacterial; D007700:Kinetics; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D010088:Oxidoreductases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D001616:beta-Galactosidase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D000911:Antibodies, Monoclonal; D003573:Cytochrome b Group; D003580:Cytochromes; D004251:DNA Transposable Elements; D045222:Electron Transport Chain Complex Proteins; D000939:Epitopes; D029968:Escherichia coli Proteins; D046911:Macromolecular Substances; D009838:Oligodeoxyribonucleotides; D011993:Recombinant Fusion Proteins; D017937:Cytochrome d Group; D010088:Oxidoreductases; C040077:cytochrome bd terminal oxidase complex, E coli; D000469:Alkaline Phosphatase; D001616:beta-Galactosidase