Isolation and characterization of the mitochondrial ATP synthase from Chlamydomonas reinhardtii. cDNA sequence and deduced protein sequence of the alpha subunit.
We have isolated the F0F1-ATP synthase complex from oligomycin-sensitive mitochondria of the green alga Chlamydomonas reinhardtii. A pure and active ATP synthase was obtained by means of sonication, extraction with dodecyl maltoside and ion exchange and gel permeation chromatography in the presence of glycerol, DTT, ATP and PMSF [corrected]. The enzyme consists of 14 subunits as judged by SDS-PAGE. A cDNA clone encoding the ATP synthase alpha subunit has been sequenced. The deduced protein sequence contains a presequence of 45 amino acids which is not present in the mature protein. The mature protein is 58-70% identical to corresponding mitochondrial proteins from other organisms. In contrast to the ATP synthase beta subunit from C. reinhardtii (Franzen and Falk, Plant Mol Biol 19 (1992) 771-780), the protein does not have a C-terminal extension. However, the N-terminal domain of the mature protein is 15-18 residues longer than in ATP synthase alpha subunits from other organisms. Southern blot analysis indicates that the protein is encoded by a single-copy gene.
Department of Biochemistry, Arrhenius laboratories, Stockholm University, Sweden.
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D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016825:Chlamydomonas reinhardtii / Q000737:chemistry / Q000201:enzymology / Q000235:genetics*; D002736:Chloroplasts / Q000201:enzymology / Q000276:immunology; D003001:Cloning, Molecular; D003429:Cross Reactions; D018076:DNA, Complementary / Q000235:genetics; D018628:Gene Dosage; D008928:Mitochondria / Q000201:enzymology / Q000235:genetics*; D008969:Molecular Sequence Data; D009840:Oligomycins / Q000494:pharmacology; D011487:Protein Conformation; D011498:Protein Precursors / Q000235:genetics; D006180:Proton-Translocating ATPases / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid; D014475:Uncoupling Agents / Q000494:pharmacology
D018076:DNA, Complementary; D009840:Oligomycins; D011498:Protein Precursors; D014475:Uncoupling Agents; D006180:Proton-Translocating ATPases