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SUMMARY: To identify components of the gliding bacterium Cytophaga johnsonae that are involved in gliding motility, we generated hybridomas that produced monoclonal antibodies (MAbs) against membranes from wild-type cells and screened for MAbs that failed to bind to cells of nonmotile mutants. Of 22 hybridomas generated, three produced MAbs that were positive in ELISA with wild-type cells or their membranes and were negative in ELISAs with 57 of 63 mutants tested. Immunoblots of polyacrylamide gels of wild-type membrane proteins showed that all three of these motility-related MAbs recognized the same antigen: a 40 kDa major membrane protein of wild-type cells. Immunoblots of nonmotile mutant cells and membranes showed that those giving negative ELISA results with the three MAbs actually produced the protein, but in only trace amounts compared with the parental strain. These results show that synthesis of the 40 kDa protein is related somehow to the ability of the cells to move, but the nature of the relationship is still unknown. The protein may be required for motility, or regulation of its synthesis and assembly may be linked to motility although it has no direct role in motility. Results of other experiments on the distribution of an immunologically related protein among other gliding bacteria and on the effects of the three motility-related MAbs on gliding of C. johnsonae did not distinguish between these two possibilities.

Monoclonal Antibodies against a Protein Missing from Nonmotile Mutants of the Gliding Bacterium Cytophaga johnsonae