ORGN 693 |
| David J. Austin, Elizabeth J. Videlock, Victor K. Chung, Michael A. Mohan, and Timothy M. Strok. Department of Chemistry, Yale University, 225 Prospect Street, New Haven, CT 06520 |
| We have recently been developing a functional protein resource for the small-molecule investigation of phosphopeptide binding interaction domains involved in protein-protein interactions. Interaction domains, as modular segments of larger proteins, have the ability to fold and function independently and are involved in a variety of signal transduction and cell regulatory mechanisms. In addition, many of these domains are known to interact with peptides and non-peptidic small molecules. Our approach is to use small-molecule probes, in conjunction with cDNA phage display, to both identify and evaluate interaction domains in the human proteome. We recently synthesized a degenerate phosphotyrosine peptide probe pYZZZ (where Z=all amino acids except tryptophan and cysteine), based on the canonical SH2-domain binding peptide motif. Screening of cDNA phage display libraries lead to the isolation of a number of SH2-domain displaying phage particles. Subsequent on-phage binding analyisis showed both dose-dependent phosphopeptide binding and phosphate-dependent binding, which is consistent with the SH2 domain phenotype. Future goals involve the isolation of additional SH2 domains from the human proteome in order to construct SH2-domain phage arrays, in a multi-well plate format, which will allow the rapid evaluation of SH2-domain binding selectivity for any phosphoprotein or probe. |
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Proteins, Peptides, Amino Acids, and Nucleotides
8:30 AM-11:30 AM, Thursday, September 11, 2003 Sheraton New York -- Royal Ballroom B, Oral
Division of Organic Chemistry |