ANALYSIS OF THE EPITOPES RECOGNIZED BY MOUSE MONOCLONAL-ANTIBODIES DIRECTED TO YERSINIA-ENTEROCOLITICA HEAT-SHOCK-PROTEIN-60

To determine amino acid sequences of the epitopes recognized by monocl onal antibodies (mAbs) 3C8 and 5C3 directed against Yersinia enterocol itica heat-shock protein (HSP60), a dot blot analysis was perfomed usi ng synthesized peptides of Y. enterocolitica HSP60 such as peptides p3 16-342, p327-359, p340-366, p316-326, p316-321, p319-323, and p321-326 which represent positions of amino acids in Y. enterocolitica HSP60. The dot blot analysis revealed that 5C3 mAb reacted with p316-342, p31 6-326 and p321-326, and 3C8 mAb p316-342 and p316-326. These results i ndicate that the epitopes recognized by the mAbs were associated with eleven amino acids, Asp Leu Gly Gin Ala Lys Arg Val Val Ile Asn, of p3 16-326. The sequence homology between p316-326 of Y. enterocolitica HS P60 and the rest of the HSP60 family suggests that the five amino acid s of Lys, Arg, Val, Ile and Asn, which are highly conserved in the HSP 60 family, might be related with the epitope recognized by 3C8. In con trast, it was also demonstrated that three amino acids of Leu, Gly and Val, which are not well conserved in the HSP60 family, might be relat ed to the epitope recognized by 5C3.