Isolation and characterization of the mitochondrial ATP synthase from Chlamydomonas reinhardtii. cDNA sequence and deduced protein sequence of the alpha subunit. [Erratum: Feb 1997, v. 33 (3), p. 571.]

• Published in: Plant molecular biology 1996-09, Vol.31 (6), p.1105-1116
• Main Authors: Nurani, G, Franzen, L.G
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; amino acid sequences; Animals; Base Sequence; Chlamydomonas reinhardtii; Chlamydomonas reinhardtii - chemistry; Chlamydomonas reinhardtii - enzymology; Chlamydomonas reinhardtii - genetics; Chloroplasts - enzymology; Chloroplasts - immunology; Cloning, Molecular; complementary DNA; Cross Reactions; DNA, Complementary - genetics; enzyme activity; Freshwater; genbank/x94149; Gene Dosage; h+-transporting atp synthase; mitochondria; Mitochondria - enzymology; Mitochondria - genetics; Molecular Sequence Data; nucleotide sequences; Oligomycins - pharmacology; Protein Conformation; Protein Precursors - genetics; Proton-Translocating ATPases - chemistry; Proton-Translocating ATPases - genetics; Proton-Translocating ATPases - immunology; purification; pyrophosphatases; Sequence Analysis; Sequence Homology, Amino Acid; structural genes; Uncoupling Agents - pharmacology; zymogens
• ISSN: 0167-4412; 1573-5028
• DOI: 10.1007/BF00040828

We have isolated the FoF1-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 -21. The enzyme consists of 14 subunits as judged by SDS-PAGE. A cDNA clone encoding the ATP synthase beta 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.