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Abstract

Methyl-coenzyme M reductase (MCR) catalyzes the reduction of methyl-coenzyme M (CH3–S–CoM) to methane. The enzyme contains as a prosthetic group the nickel porphinoid F430 which in the active enzyme is in the EPR-detectable Ni(I) oxidation state. Crystal structures of several inactive Ni(II) forms of the enzyme but not of the active Ni(I) form have been reported. To obtain structural information on the active enzyme–substrate complex we have now acquired X-ray absorption spectra of active MCR in the presence of either CH3–S–CoM or the substrate analog coenzyme M (HS–CoM). For both MCR complexes the results are indicative of the presence of a five-coordinate Ni(I), the five ligands assigned as four nitrogen ligands from F430 and one oxygen ligand. Analysis of the spectra did not require the presence of a sulfur ligand indicating that CH3–S–CoM and HS–CoM were not coordinated via their sulfur atom to nickel in detectable amounts. As a control, X-ray absorption spectra were evaluated of three enzymatically inactive MCR forms, MCR-silent, MCR-ox1-silent and MCR-ox1, in which the nickel is known to be six-coordinate. Comparison of the edge position of the X-ray absorption spectra revealed that the Ni(I) in the active enzyme is more reduced than the Ni in the two EPR-silent Ni(II) states. Surprisingly, the edge position of the EPR-active MCR-ox1 state was found to be the same as that of the two silent states indicating similar electron density on the nickel. Electronic supplementary material is available if you access this article at http://www.dx.doi.org/10.1007/s00775-002-0399-2. On that page (frame on the left side), a link takes you directly to the supplementary material.