Frank Otto Gombert - Pentapeptide identified as a monoclonal antibody binding site in the serine-protease domain of t-PA

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  Publication Details (including relevant citation   information):

  Gombert,   F. O.; Werz, W.; Schlüter, M.; Bayer, A.; Werner, R. G.;   Berthold, W.; Jung, G. Pentapeptide identified as a monoclonal   antibody binding site in the serine-protease domain of t-PA.   Biological chemistry Hoppe-Seyler 1994,  375, 471.


  The first defined sequential epitope of the tissue plasminogen   activator (t-PA) was determined by a monoclonal antibody against   a synthetic peptide segment corresponding to peptide sequence   341-354 of t-PA. This segment was selected by computer assisted   epitope prediction. Balb/c mice were immunized with   catalase-peptide and   tripalmitoyl-S-glyceryl-cys-teinyl-seryl-peptide conjugates. A   monoclonal antibody derived from this immunization was reactive   with native recombinant t-PA (rt-PA) and reduced   carboxymethylated recombinant t-PA (RCM rt-PA). The sequential   epitope was detected by Pepscan method using overlapping octa-   and nonapeptides. By fine epitope mapping with tetra-, penta-,   hexa- and heptapeptides the epitope was minimized to the   pentapeptide EEEQK (347-351). Replacement set analysis confirmed   the importance of this amino acid sequence, especially of the   amino acid E348, for antibody binding. Functional assays of rt-PA   were not affected by this antibody indicating that the epitope   has no influence on the enzymatic center and the binding site of   the inhibitor. The analysis demonstrates that the predicted   recognition site of the monoclonal antibody 17-134/11 is exposed   on the surface of the native rt-PA molecule

  Address (URL): 3.1994.375.7.471.xml