Figure 2.

Mdm2 encompasses two putative 2A10 epitopes. (A) A 6-mer phage display peptide library was screened with the 2A10 antibody as described in Materials and Methods. Eleven positive clones obtained after the second panning, as well as four positive clones obtained after the third panning, were subjected to DNA sequencing. All clones were found to fall into only two sequence classes. The deduced amino acid sequences of the corresponding 6-mer peptides, present in the positive phage, are shown. Based on this data and on the screening of another independent library (data not shown), the consensus 2A10 epitope was deduced as 1-DYS-5, where 1 is preferentially D and 5 is preferentially L; the core DYS sequence is indicated in bold. Also shown is the number of positive phage found to carry the indicated sequence, out the total number of positive phage (in parentheses) sequenced after each panning. (B) Positions of the two putative 2A10 epitopes on Mdm2 are indicated. The positions of the main identified structural domains of the protein (Momand et al. 2000) are also indicated. Zn finger = zinc finger.