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Isolation and characterization of the mitochondrial ATP synthase fromChlamydomonas reinhardtii. cDNA sequence and deduced protein sequence of the α subunit

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Abstract

We have isolated the F0F1-ATP synthase complex from oligomycin-sensitive mitochondria of the green algaChlamydomonas reinhardtii. A pure and active ATP synthase was obtained by eans 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 α 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 β subunit fromC. 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 α subunits from other organisms. Southern blot analysis indicates that the protein is encoded by a single-copy gene.

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Abbreviations

DM:

dodecyl-β-D-maltoside

OSCP:

oligomycin sensitivity conferring protein

PMSF:

phenyl-methylsulfonylfluoride

DTT:

dithiothreitol

EDTA:

ethylenediaminotetraacetic disodium salt

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Authors and Affiliations

  1. Department of Biochemistry Arrhenius laboratories, Stockholm University, S-106 91, Stockholm, Sweden

    Ghasem Nurani

  2. Department of Plant Physiology, Botanical Institute, Göteborg University, Carl Skottsbergs gata 22B, S-413 19, Göteborg, Sweden

    Lars-Gunnar Franzén

Authors
  1. Ghasem Nurani
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  2. Lars-Gunnar Franzén
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Nurani, G., Franzén, LG. Isolation and characterization of the mitochondrial ATP synthase fromChlamydomonas reinhardtii. cDNA sequence and deduced protein sequence of the α subunit. Plant Mol Biol 31, 1105–1116 (1996). https://doi.org/10.1007/BF00040828

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  • DOI: https://doi.org/10.1007/BF00040828

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