The studies described here were performed to characterize further the plasma membrane associated protein BsSco, which is the product of the gene ypmQ, in Bacillus subtilis. BsSco is a member of the Sco family of proteins found in the inner mitochondrial membrane of yeast and humans and implicated as an accessory protein in the assembly of the Cu-A site of cytochrome c oxidase. We have cloned the gene expressing BsSco, placed a six-histidine tag on its C-terminus, and over-expressed this protein in B. subtilis. Recombinant BsSco with the his-tag has been purified from Triton X-100-solubilized plasma membranes by nickel metal affinity chromatography. Mass spectral analysis of the purified protein is consistent with processing of BsSco by signal peptidase 11 removing an N-terminal putative transmembrane sequence to leave an acyl-glyceryl moiety at cysteine residue 19. Antibodies, raised against purified, recombinant BsSco, were used to characterize the timing of the level of native BsSco in batch cultures of wild-type R subtilis. There is a marked lag in the level of native BsSco, but it does appear prior to cytochrome c oxidase, which is expressed in late stage growth. This work supports a role for BsSco in the assembly of the Cu-A site of cytochrome C oxidise and its functional relationship to the Sco proteins found in eukaryotic cells. (C) 2003 Elsevier Inc. All rights reserved.