PMID: 9654068Jul 8, 1998Paper

Methanol:coenzyme M methyltransferase from Methanosarcina barkeri--identification of the active-site histidine in the corrinoid-harboring subunit MtaC by site-directed mutagenesis

European Journal of Biochemistry
K Sauer, R K Thauer

Abstract

The enzyme system catalyzing the formation of methyl-coenzyme M from methanol and coenzyme M in Methanosarcina barkeri is composed of the three different polypeptides MtaA, MtaB and MtaC of which MtaC harbors a corrinoid prosthetic group. The heterologous expression of mtaA and mtaB in Escherichia coli has been described previously. We report here on the overproduction of the apoprotein of MtaC in E. coli, on its reconstitution to the active holoprotein with either cob(II)alamin or methyl-cob(III)alamin, and on the properties of the reconstituted corrinoid protein. Reconstituted MtaC was found to contain 1 mol bound cobamide/mol. EPR spectroscopic evidence is presented for a His residue as an axial ligand to Co2+ of the bound corrinoid. This active-site His was identified by site-directed mutagenesis as His136 in the MtaC sequence that contains four His residues. The reconstituted MtaC, in the cob(I)amide oxidation state, was methylated with methanol in the presence of MtaB and demethylated with coenzyme M in the presence of MtaA. In the presence of both MtaB and MtaA, methyl-coenzyme M was formed from methanol and coenzyme M at specific rates comparable to those determined for the enzyme system purified from M. barkeri. M. bar...Continue Reading

Citations

Apr 28, 2000·European Journal of Biochemistry·K Sauer, R K Thauer
Dec 5, 2006·Proceedings of the National Academy of Sciences of the United States of America·Christoph H HagemeierUlrich Ermler
Mar 3, 2006·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Weihong ZhouBi-Cheng Wang
Dec 5, 2008·Archaea : an International Microbiological Journal·Adam M GussWilliam W Metcalf
Dec 9, 2008·Current Opinion in Structural Biology·Rowena G MatthewsSupratim Datta
Dec 20, 2005·Molecular Microbiology·Anirban MahapatraJoseph A Krzycki
Mar 17, 1999·FEMS Microbiology Reviews·J G Ferry
Feb 10, 2021·Current Opinion in Microbiology·Katie E Shalvarjian, Dipti D Nayak
Jun 9, 2005·Chemical Reviews·Kenneth L Brown
Apr 4, 2015·The Journal of Physical Chemistry. a·Piotr LodowskiPawel M Kozlowski

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