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Purification of component C from Methanosarcina mazei and immunolocalization in Methanosarcinaceae

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Abstract

Studies on immunological relationships among Methanosarcina genus using immunofluorescence and immunoprecipitation showed that a common antigen can be extracted by shaking in aqueous phase. This antigen was purified from Methanosarcina mazei. The protein had a molecular weight of 283400 daltons with three subunits, α=68000, β=43200 and γ=30500. It contained nickel, coenzyme M and F430. Its biochemical characteristics identified this antigen as the component C of the methyl CoM reductase complex. But EPR study showed that the nickel was Ni(II). Biological activity was detectable neither by heterologous in vitro assay nor by the DTT assay. Immunogold labelling showed that the component C was located randomly in the cytoplasm in Methanosarcina species and in Methanothrix soehngenii. In addition, specific labelling was also observed outside of the heteropolysaccharidic envelopes probably due to the absorption of component C released by the lysis of some cells in the clumps.

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Authors and Affiliations

  1. Station de Technologie Alimentaire, Institut National de la Recherche Agronomique, BP 39, F-59651, Villeneuve d'Ascq Cedex, France

    I. Thomas, H.-C. Dubourguier, P. Debeire & G. Albagnac

  2. Institut National de la Santé et de la Recherche Médicale, U 42, CERTIA, BP 39, F-59651, Villeneuve d'Ascq Cedex, France

    G. Prensier

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  1. I. Thomas
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  2. H.-C. Dubourguier
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  3. G. Prensier
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  4. P. Debeire
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  5. G. Albagnac
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Thomas, I., Dubourguier, HC., Prensier, G. et al. Purification of component C from Methanosarcina mazei and immunolocalization in Methanosarcinaceae . Arch. Microbiol. 148, 193–201 (1987). https://doi.org/10.1007/BF00414811

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  • DOI: https://doi.org/10.1007/BF00414811

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