Molybdenum requirement for translocation of dimethyl sulfoxide reductase to the periplasmic space in a photodenitrifier, Rhodobacter sphaeroides f. sp. denitrificans.
Translocation of dimethyl sulfoxide (DMSO) reductase to the periplasmic space was studied in vivo with a photodenitrifier, Rhodobacter sphaeroides f. sp. denitrificans, using immunoblotting analysis and radioactive labeling. A polypeptide with an apparent molecular mass about 2,000 Da higher than that of DMSO reductase accumulated during induction of the reductase with DMSO. An uncoupler, carbonyl cyanide-m-chlorophenylhydrazone, inhibited the processing of the polypeptide after cells had been radioactively pulse-labeled with [35S]methionine. These results indicated that the higher-molecular-mass polypeptide was the precursor form of DMSO reductase. The precursor form accumulated in either the cytoplasm or the membrane, whereas the mature form accumulated in the periplasmic space. The membrane-bound precursor was sensitive to proteinase K treatment from both the cytoplasmic and periplasmic sides of the membrane, indicating that the polypeptide binds to the membrane, exposing it to both the outer and inner surfaces of the cytoplasmic membrane. Processing of the precursor was hampered by removal of molybdate from the medium and was restored by its readdition. It was also inhibited by the addition of tungstate in the medium.
Yoshida
Y
Y
Department of Biology, Tokyo Metropolitan University, Japan.
Takai
M
M
Satoh
T
T
Takami
S
S
D001692:Biological Transport / Q000187:drug effects; D015153:Blotting, Western; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone / Q000494:pharmacology; D002462:Cell Membrane / Q000502:physiology; D004591:Electrophoresis, Polyacrylamide Gel; D019286:Endopeptidase K; D007506:Iron-Sulfur Proteins; D008982:Molybdenum / Q000494:pharmacology*; D010088:Oxidoreductases / Q000493:pharmacokinetics*; D010626:Phenylethyl Alcohol / Q000494:pharmacology; D011499:Protein Processing, Post-Translational / Q000187:drug effects / Q000502:physiology; D012242:Rhodobacter sphaeroides; D012697:Serine Endopeptidases / Q000494:pharmacology
D007506:Iron-Sulfur Proteins; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone; D008982:Molybdenum; D010088:Oxidoreductases; C045664:dimethyl sulfoxide reductase; D012697:Serine Endopeptidases; D019286:Endopeptidase K; D010626:Phenylethyl Alcohol