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J. Biol. Chem., Vol. 279, Issue 10, 8802-8807, March 5, 2004

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An Oriented Peptide Array Library (OPAL) Strategy to Study Protein-Protein Interactions^*

Maria Rodriguez, Shawn S.-C. Li, J. Wade Harper¶, and Zhou Songyang||

From the Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, Texas 77030 and the Department of Biochemistry, Faculty of Medicine and Dentistry, University of Western Ontario, London, Ontario N6A 5C1, Canada

One of the major questions in signal transduction is how the specificities of protein-protein interactions determine the assembly of distinct signaling complexes in response to stimuli. Several peptide library methods have been developed and widely used to study proteinprotein interactions. These approaches primarily rely on peptide or DNA sequencing to identify the peptide or consensus motif for binding and may prove too costly or difficult to accommodate high throughput applications. We report here an oriented peptide array library (OPAL) approach that should facilitate high throughput proteomic analysis of protein-protein interactions. OPAL integrates the principles of both the oriented peptide libraries and array technologies. Hundreds of pools of oriented peptide libraries are synthesized as amino acid scan arrays. We demonstrate that these arrays can be used to map the specificities of a variety of interactions, including antibodies, protein domains such Src homology 2 domains, and protein kinases.

Received for publication, October 29, 2003 , and in revised form, December 15, 2003.

^* This work was supported by National Institutes of Health Grants CA84208 (to Z. S.) and AG11085 (to J. W. H.), Initiative for Minority Student Development Grant GM569209 (to M. R.), and grants from the Welch Foundation (to Z. S.) and the Specialized Program of Research Excellence (SPORE) in Prostate Cancer (to J. W. H.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^¶ Present address: Dept. of Pathology, Harvard Medical School, Boston, MA 02115.

^|| To whom correspondence should be addressed. Tel.: 713-798-5220; Fax: 713-796-9438; E-mail: songyang{at}bcm.tmc.edu.

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