Localization and solubilization of the respiratory nitrate reductase of Bacillus stearothermophilus.
Membranes were isolated from Bacillus stearothermophilus 2184D by lysozyme digestion of the cell wall and subsequent differential centrifugation. Observations with the electron microscope indicate that such membranes are relatively intact and have a typical membrane appearance. Nitrate will preferentially oxidize the cytochrome b of such membranes. Approximately 80% of the total respiratory nitrate reductase activity of whole cells can be localized in the washed membrane fraction and the process of membrane isolation results in a sixfold purification of this enzyme. Of several detergents tested, sodium dodecyl sulfate, Triton 114, and Triton X-100 are most effective in converting reduced methyl viologen-nitrate reductase to a form which will not pellet at 130,000 x g. Density gradient analysis reveals that such detergent-mediated solubilization converts virtually all membrane protein to a form of lighter density.
D000085:Acetates; D001407:Bacillus / Q000166:cytology / Q000201:enzymology* / Q000254:growth & development; D002462:Cell Membrane / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D002473:Cell Wall / Q000187:drug effects; D002499:Centrifugation, Density Gradient; D003470:Culture Media; D003580:Cytochromes / Q000378:metabolism; D003902:Detergents / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D008854:Microscopy, Electron; D009113:Muramidase / Q000494:pharmacology; D009243:NAD; D009566:Nitrates / Q000378:metabolism; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000302:isolation & purification / Q000378:metabolism; D012995:Solubility; D013395:Sucrose; D014501:Uranium
D000085:Acetates; D003470:Culture Media; D003580:Cytochromes; D003902:Detergents; D007202:Indicators and Reagents; D009566:Nitrates; D009243:NAD; D014501:Uranium; D013395:Sucrose; D010088:Oxidoreductases; D009113:Muramidase