Identification of the mcrD gene product and its association with component C of methyl coenzyme M reductase in Methanococcus vannielii.

A mcrD-lacZ gene fusion has been constructed and expressed under lacP control in Escherichia coli. Antibodies raised against the product of this gene fusion have been used in Western blotting (immunoblotting) to demonstrate the gene product of mcrD (gpmcrD) in Methanococcus vannielii. The alpha, beta, and gamma subunit polypeptides of component C of methyl coenzyme M reductase (MR) were coprecipitated with gpmcrD when bound by antibodies raised either against MR or against gpmcrD-lacZ. This association of MR and gpmcrD did not withstand polyacrylamide gel electrophoresis under nondenaturing conditions.