Biosynthesis and membrane binding of succinate dehydrogenase in Bacillus subtilis.
Antibodies specific for the Mr 65,000 (flavoprotein) and the Mr 28,000 subunits of the succinic dehydrogenase (SDH) of Bacillus subtilis were obtained. By using these antibodies it was shown that both subunits accumulated in the cytoplasm during 5-aminolevulinic acid starvation of a 5-aminolevulinic acid auxotroph. In the cytoplasm the subunits were not associated since they precipitated essentially independently of each other with subunit-specific antibody. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis the cytoplasmic subunits migrated identically with the corresponding subunits from the purified membrane-bound SDH complex. Cytoplasmic subunits were pulse-labeled with L-[35S]methionine during 5-aminolevulinic acid starvation. The labeled subunits bound to the membrane when heme synthesis was resumed and also when protein synthesis was blocked by chloramphenicol before readdition of 5-aminolevulinic acid. The experiments thus demonstrated a precursor relationship between cytoplasmic subunits and the subunits of the membrane-bound SDH complex. All SDH-negative mutants isolated so far carry mutations in the citF locus. None of the mutants was found to have either the Mr 65,000 or the Mr 28,000 SDH subunits in the membrane. Four citF mutants, however, contained both subunits in the cytoplasm. Three of these mutants lacked spectrally detectable cytochrome b558. The respective mutations mapped at one end of the citF locus. These results strongly support our previous suggestion that cytochrome b558 is (part of) a membrane binding site for SDH in B. subtilis.
D000622:Aminolevulinic Acid / Q000378:metabolism; D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics; D002462:Cell Membrane / Q000201:enzymology; D003573:Cytochrome b Group; D003580:Cytochromes / Q000378:metabolism; D003593:Cytoplasm / Q000201:enzymology; D009154:Mutation; D013385:Succinate Dehydrogenase / Q000378:metabolism*
D003573:Cytochrome b Group; D003580:Cytochromes; D000622:Aminolevulinic Acid; D013385:Succinate Dehydrogenase