Antigenic properties of the GroEL‐like protein of Campylobacter rectus

The purpose of this study was to clarify the antigenic properties of the GroEL-like protein of Campylobacter rectus using a specific polyclonal antibody directed to the purified 64-kDa GroEL-like protein (pAb-CrGroEL), a polyclonal antibody directed to the Actinobacillus actinomycetemcomitans GroEL-like protein (pAb-AaGroEL) and a monoclonal antibody against the recombinant human HSP60 (mAb-HuHSP60). In SDS-PAGE/Western immunoblotting analysis, mAb-HuHSP60, pAb-CrGroEL and pAb-AaGroEL were found to react with the GroEL-like protein (64-kDa) present in all C. rectus strains. A 150-kDa protein in C. rectus ATCC 33238 also reacted strongly with pAb-CrGroEL. This 150-kDa protein was found to be present on the surface-associated material of bacterial cells, as determined by transmission electron microscopy and immunogold labelling of cells with pAb-CrGroEL. Analysis of the first 20 N-terminal amino acids of the sequence of the 150-kDa protein revealed a strong homology (80%) with the C. rectus surface layer (S-layer) protein. Investigation of the biochemical nature of antigenic determinants using periodic acid and proteolytic enzymes showed that the C. rectus GroEL-like protein possessed immunodominant epitopes in both peptide and carbohydrate chains, and that the immunoreactive determinants of the 150-kDa protein belonged to carbohydrate. These results suggest that the GroEL-like protein and the S-layer protein of C. rectus may share the same carbohydrate epitopes.