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Identification of a novel nifH-like (frxC) protein in chloroplasts of the liverwort Marchantia polymorpha

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Abstract

The frxC gene, one of the unidentified open reading frames present in liverwort chloroplast DNA, shows significant homology with the nifH genes coding for the Fe protein, a component of the nitrogenase complex (Ohyama et al., 1986, Nature 322: 572–574). A truncated form of the frxC gene was designed to be over-expressed in Escherichia coli and an antibody against this protein was prepared using the purified product as an antigen. This antibody reacted with a protein in the soluble fraction of liverwort chloroplasts, which had an apparent molecular weight of 31 000, as revealed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, in good agreement with a putative molecular weight of 31945 deduced from the DNA sequence of the frxC gene. In a competitive inhibition experiment, the antigenicity of this protein was indicated to be similar to that of the over-expressed protein in E. coli. Therefore, we concluded that the frxC gene was expressed in liverwort chloroplasts and that its product existed in a soluble form. The molecular weight of the frxC protein was approximately 67 000, as estimated by gel filtration chromatography, indicating that the frxC protein may exist as a dimer of two identical polypeptides analogous to the Fe protein of nitrogenase. The results obtained from affinity chromatography supported the possibility that the frxC protein, which possesses a ATP-binding sequence in its N-terminal region that is conserved among various other ATP-binding proteins, has the ability to bind ATP.

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Authors and Affiliations

  1. Department of Biology, Faculty of Science, Osaka University, 560, Toyonaka, Osaka, Japan

    Yuichi Fujita, Yasuhiro Takahashi & Hiroshi Matsubara

  2. Research Center for Cell and Tissue Culture, Faculty of Agriculture, Kyoto University, 606, Kyoto, Japan

    Takayuki Kohchi & Kanji Ohyama

  3. Department of Biophysics, Faculty of Science, Kyoto University, 606, Kyoto, Japan

    Haruo Ozeki

Authors
  1. Yuichi Fujita
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  2. Yasuhiro Takahashi
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  3. Takayuki Kohchi
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  4. Haruo Ozeki
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  5. Kanji Ohyama
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  6. Hiroshi Matsubara
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Fujita, Y., Takahashi, Y., Kohchi, T. et al. Identification of a novel nifH-like (frxC) protein in chloroplasts of the liverwort Marchantia polymorpha . Plant Mol Biol 13, 551–561 (1989). https://doi.org/10.1007/BF00027315

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  • DOI: https://doi.org/10.1007/BF00027315

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