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  Molecular characterization of methanogenic N5-methyltetrahydromethanopterin: Coenzyme M methyltransferase

Upadhyay, V., Ceh, K., Tumulka, F., Abele, R., Hoffmann, J., Langer, J. D., et al. (2016). Molecular characterization of methanogenic N5-methyltetrahydromethanopterin: Coenzyme M methyltransferase. Biochimica et Biophysica Acta-Biomembranes, 1858(9), 2140-2144. doi:10.1016/j.bbamem.2016.06.011.

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 Creators:
Upadhyay, Vikrant1, Author              
Ceh, Katharina1, Author              
Tumulka, Franz2, Author
Abele, Rupert2, Author
Hoffmann , Jan3, Author
Langer, Julian D.1, Author              
Shima, Seigo3, 4, Author              
Ermler, Ulrich1, Author              
Affiliations:
1Department of Molecular Membrane Biology, Max Planck Institute of Biophysics, Max Planck Society, ou_2068290              
2Goethe-Universität Frankfurt am Main, Institut für Biochemie, Max-von-Laue-Straße 9, 60438 Frankfurt am Main, Germany, ou_persistent22              
3Max-Planck-Institut für terrestrische Mikrobiologie, Karl-von-Frisch-Straße, D-35043 Marburg, Germany, ou_persistent22              
4PRESTO, Japan Science and Technology Agency (JST), Honcho, Kawaguchi, Saitama 332-0012, Japan, ou_persistent22              

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Free keywords: Methanogenesis N5-methyl-tetrahydromethanopterin; coenzyme M methyltransferase; Membrane protein complex; SEC-MALS; LILBID-MS; Dimethyl maleic anhydride
 Abstract: Methanogenic archaea share one ion gradient forming reaction in their energy metabolism catalyzed by the membrane-spanning multisubunit complex N5-methyl-tetrahydromethanopterin: coenzyme M methyltransferase (MtrABCDEFGH or simply Mtr). In this reaction the methyl group transfer from methyl-tetrahydromethanopterin to coenzyme M mediated by cobalamin is coupled with the vectorial translocation of Na+across the cytoplasmic membrane. No detailed structural and mechanistic data are reported about this process. In the present work we describe a procedure to provide a highly pure and homogenous Mtr complex on the basis of a selective removal of the only soluble subunit MtrH with the membrane perturbing agent dimethyl maleic anhydride and a subsequent two-step chromatographic purification. A molecular mass determination of the Mtr complex by laser induced liquid bead ion desorption mass spectrometry (LILBID-MS) and size exclusion chromatography coupled with multi-angle light scattering (SEC-MALS) resulted in a (MtrABCDEFG)3 heterotrimeric complex of ca. 430 kDa with both techniques. Taking into account that the membrane protein complex contains various firmly bound small molecules, predominantly detergent molecules, the stoichiometry of the subunits is most likely 1:1. A schematic model for the subunit arrangement within the MtrABCDEFG protomer was deduced from the mass of Mtr subcomplexes obtained by harsh IR-laser LILBID-MS.

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Language(s): eng - English
 Dates: 2016-05-042016-06-152016-06-212016-09
 Publication Status: Published in print
 Pages: 5
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1016/j.bbamem.2016.06.011
 Degree: -

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Title: Biochimica et Biophysica Acta-Biomembranes
Source Genre: Journal
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Publ. Info: Amsterdam : Elsevier
Pages: - Volume / Issue: 1858 (9) Sequence Number: - Start / End Page: 2140 - 2144 Identifier: ISSN: 0005-2736
CoNE: https://pure.mpg.de/cone/journals/resource/954926938702