Studying receptor-ligand interactions using encoded amino acid scanning

Julio A. Camarero, Brenda Ayers, Tom W. Muir

Research output: Contribution to journalArticlepeer-review

8 Scopus citations


A novel technique is described that allows the synthesis, functional analysis, and quantitative readout of defined arrays of polypeptide analogues in aqueous solution. Key to this approach is the use of a simple encoding- decoding system in which a unique Fmoc-amino acid tag is covalently attached to the C terminus of each member of a molecular array through a selectively cleavable bond. These tags can be cleanly removed from the molecules they encode, allowing single-step characterization and quantification of the entire mixture by HPLC. The utility of this technique is illustrated through the preparation of an array of proline-rich sequences based on the exchange factor C3G, one of the natural ligands of the N-terminal SH3 domain from the proto-oncogene, c-Crk. The array was designed to systematically modify those residues within the C3G peptide ligand thought to make key interactions with the c-Crk SH3 domain. Using competition binding experiments, it was possible to determine the relative ED50 values for the entire array of molecules simultaneously. These studies revealed that in order to maintain optimal binding to the SH3 domain, the P-3 side chain of the ligand must be positively charged and the P-0 side chain must be hydrophobic and extend beyond the γ-carbon. The excellent correlation between these relative ED50 values and a series of relative K(d) values determined from individual peptides suggests that this approach may be useful in determining, in a parallel fashion, the relative biological activities of arrays of polypeptides.

Original languageEnglish (US)
Pages (from-to)7487-7495
Number of pages9
Issue number20
StatePublished - May 19 1998
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biochemistry


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