Methanogenesis marker protein 10 (Mmp10) from methanosarcina acetivorans is a radical S-adenosylmethionine methylase that unexpectedly requires cobalamin

Matthew I. Radle, Danielle V. Miller, Tatiana N. Laremore, Squire J. Booker

Research output: Contribution to journalReview article

6 Scopus citations

Abstract

Methyl coenzyme M reductase (MCR) catalyzes the last step in the biological production of methane by methanogenic archaea, as well as the first step in the anaerobic oxidation of methane to methanol by methanotrophic archaea. MCR contains a number of unique post-translational modifications in its subunit, including thioglycine, 1-N-methylhistidine, S-methylcysteine, 5-C-(S)-methylarginine, and 2-C-(S)-methyl-glutamine. Recently, genes responsible for the thioglycine and methylarginine modifications have been identified in bioinformatics studies and in vivo complementation of select mutants; however, none of these reactions has been verified in vitro. Herein, we purified and biochemically characterized the radical S-adenosylmethionine (SAM) protein MaMmp10, the product of the methanogenesis marker protein 10 gene in the methane-producing archaea Methanosarcina acetivorans. Using an array of approaches, including kinetic assays, LC-MS–based quantification, and MALDI TOF–TOF MS analyses, we found that MaMmp10 catalyzes the methylation of the equivalent of Arg285 in a peptide substrate surrogate, but only in the presence of cobalamin. We noted that the methyl group derives from SAM, with cobalamin acting as an intermediate carrier, and that MaMmp10 contains a C-terminal cobalamin-binding domain. Given that Mmp10 has not been annotated as a cobalamin-binding protein, these findings suggest that cobalamin-dependent radical SAM proteins are more prevalent than previously thought.

Original languageEnglish (US)
Pages (from-to)11712-11725
Number of pages14
JournalJournal of Biological Chemistry
Volume294
Issue number31
DOIs
StatePublished - Aug 2 2019

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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