The organization of formate dehydrogenase in the cytoplasmic membrane of Escherichia coli.
The arrangement of the proton-translocating formate dehydrogenase of the anaerobic respiratory chain of Escherichia coli within the cytoplasmic membrane was examined by direct covalent modification with non-membrane-permeant reagents. Three methods were employed, lactoperoxidase-catalysed radioiodination, labelling with diazotized [125I] di-iodosulphanilic acid and labelling with diazobenzene [35S] sulphonate. All three procedures yield consistent with the view that the two larger subunits of the enzyme, Mr 110000 and 32000, both occupy transmembranous locations within the membrane. In each case the modification of the Ca2+ or Mg2+-activated F1-ATPase was monitored, and all reagents employed correctly located this enzyme at the cytoplasmic face of the membrane. A procedure involving agglutination with specific antibodies is described which appears to fractionate membrane vesicles of mixed orientation into two populations, one with the same membrane orientation as that of spheroplasts and the other opposite orientation.
D000445:Aldehyde Oxidoreductases / Q000378:metabolism*; D003593:Cytoplasm / Q000201:enzymology; D003979:Diazonium Compounds; D004926:Escherichia coli / Q000201:enzymology*; D005560:Formate Dehydrogenases / Q000276:immunology / Q000378:metabolism*; D007106:Immune Sera; D007202:Indicators and Reagents; D007425:Intracellular Membranes / Q000201:enzymology; D007457:Iodine Radioisotopes; D007784:Lactoperoxidase; D013425:Sulfanilic Acids / Q000031:analogs & derivatives
D003979:Diazonium Compounds; D007106:Immune Sera; D007202:Indicators and Reagents; D007457:Iodine Radioisotopes; D013425:Sulfanilic Acids; C007308:diazodiiodosulfanilic acid; D007784:Lactoperoxidase; D005560:Formate Dehydrogenases; D000445:Aldehyde Oxidoreductases; C020644:Formate dehydrogenase (cytochrome); C000469:diazobenzenesulfonic acid