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Significant advances have been made in the past year in our understanding of the structure, function, and mode of regulation and assembly of nickel-containing enzymes. The highlight of 1997 was the elucidation of the methyl-CoM reductase structure.

The yellow chromophore of the methyl-coenzyme M methylreductase of Methanobacterium thermoautotrophicum has been found to be the nickel-containing factor F430. Treatment of 63Ni-labeled methylreductase with 80% aqueous methanol released the radiolabel as well as the yellow chromophore; both properties were associated with a single compound that was found to be identical to F430, the stoichiometry being 1 mol of nickel per mol of F430 and 2 mol of F430 per mol of methylreductase.

The subunits and the nickel tetrapyrrole factor F430 of the methylreductase component C from Methanobacterium thermoautotrophicum have been separated from one another and reassociated to form an intact, active enzyme. The individual subunits were extracted from polyacrylamide gels after NaDodSO4/polyacrylamide gel electrophoresis and reassociated with F430 under a H2 atmosphere at 55 degrees C. When these components were reassociated in the presence of the substrate 2-(methylthio)ethanesulfonic acid, 72% of the original activity was recovered. When F430 was omitted, no activity was detected in the reconstituted methylreductase reaction. Individual subunits were inactive, when incubated in the presence of factor F430, 2-(methylthio)ethanesulfonic acid, magnesium acetate, and ATP. Evidence suggests that F430 binds to the large Mr 68,000 subunit of component C.