Methyl-coenzyme M reductase from methanogenic archaea: Isotope effects on the formation and anaerobic oxidation of methane

Silvan Scheller, Meike Goenrich, Rudolf K. Thauer, Bernhard Jaun*

*Corresponding author for this work

Research output: Contribution to journalArticleScientificpeer-review

32 Citations (Scopus)

Abstract

The nickel enzyme methyl-coenzyme M reductase (MCR) catalyzes two important transformations in the global carbon cycle: methane formation and its reverse, the anaerobic oxidation of methane. MCR uses the methyl thioether methyl-coenzyme M (CH3-S-CH2CH2-SO 3-, Me-S-CoM) and the thiol coenzyme B (CoB-SH) as substrates and converts them reversibly to methane and the corresponding heterodisulfide (CoB-S-S-CoM). The catalytic mechanism is still unknown. Here, we present isotope effects for this reaction in both directions, catalyzed by the enzyme isolated from Methanothermobacter marburgensis. For methane formation, a carbon isotope effect (12CH3-S-CoM/ 13CH3-S-CoM) of 1.04 ± 0.01 was measured, showing that breaking of the C-S bond in the substrate Me-S-CoM is the rate-limiting step. A secondary isotope effect of 1.19 ± 0.01 per D in the methyl group of CD3-S-CoM indicates a geometric change of the methyl group from tetrahedral to trigonal planar upon going to the transition state of the rate-limiting step. This finding is consistent with an almost free methyl radical in the highest transition state. Methane activation proceeds with a primary isotope effect of 2.44 ± 0.22 for the C-H vs C-D bond breakage and a secondary isotope effect corresponding to 1.17 ± 0.05 per D. These values are consistent with isotope effects reported for oxidative cleavage/reductive coupling occurring at transition metal centers during C-H activation but are also in the range expected for the radical substitution mechanism proposed by Siegbahn et al. The isotope effects presented here constitute boundary conditions for any suggested or calculated mechanism.

Original languageEnglish
Pages (from-to)14975-14984
Number of pages10
JournalJournal of the American Chemical Society
Volume135
Issue number40
DOIs
Publication statusPublished - 2013
MoE publication typeA1 Journal article-refereed

Fingerprint Dive into the research topics of 'Methyl-coenzyme M reductase from methanogenic archaea: Isotope effects on the formation and anaerobic oxidation of methane'. Together they form a unique fingerprint.

Cite this