Subunit IV (Mr = 14,384) of the cytochrome b-c1 complex from Rhodobacter sphaeroides. Cloning, DNA sequencing, and ubiquinone binding domain.
The Rhodobacter sphaeroides gene encoding subunit IV of the cytochrome b-c1 complex (fbcQ) was cloned and sequenced. The fbcQ cistron is 372 base pairs long and encodes 124 amino acid residues. The molecular mass of subunit IV, deduced from the nucleotide sequence, is 14,384 Da. A hydropathy plot of the predicted amino acid sequence revealed only one transmembrane helix; it is near the C-terminal end. The 3-azido-2-methyl-5-methoxy-6-(3,7-dimethyl[3H]octyl)-1,4-benzoquinone ([3H]azido-Q)-labeled subunit IV was isolated from the [3H]-azido-Q-treated cytochrome b-c1 complex. A ubiquinone-binding peptide was obtained by digesting the labeled subunit IV with V8 protease followed by high performance liquid chromatography separation. Amino acid analysis and partial N-terminal sequencing of this ubiquinone-binding peptide revealed that it corresponded to residues 77-124 of subunit IV. Based on the hydropathy profile and predicted tendency to form alpha-helices and beta-sheets, we propose a structural model for subunit IV. In this model the ubiquinone-binding domain is located near the surface of the membrane.
Usui
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Department of Biochemistry, Oklahoma State University, Stillwater 74078.
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D000345:Affinity Labels; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001386:Azides / Q000737:chemistry / Q000302:isolation & purification; D010582:Bacteriophage lambda / Q000235:genetics; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D003062:Codon; D004269:DNA, Bacterial / Q000235:genetics; D014450:Electron Transport Complex III / Q000235:genetics* / Q000378:metabolism; D005814:Genes, Viral; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D011487:Protein Conformation; D015183:Restriction Mapping; D012242:Rhodobacter sphaeroides / Q000201:enzymology*; D013329:Structure-Activity Relationship; D014357:Trypsin; D014451:Ubiquinone / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism* / Q000031:analogs & derivatives
D000345:Affinity Labels; D000596:Amino Acids; D001386:Azides; D003062:Codon; D004269:DNA, Bacterial; D014451:Ubiquinone; C048123:3-azido-2-methyl-5-methoxy-6-(3,7-dimethyloctyl)-1,4-benzoquinone; D014357:Trypsin; D014450:Electron Transport Complex III