Vol. 12, Issue 11, 1785-1791, November 2002

METHODS
Detection of Peptides, Proteins, and Drugs That Selectively Interact With Protein Targets

¹ Division of Basic Science, Fox Chase Cancer Center, Philadelphia, Pennsylvania 19111, USA; ² Department of Molecular Biology and Medical Biotechnology, Russian State Medical University, Moscow, Russia; ³ Dana-Farber Cancer Institute, Boston, Massachusetts 02115, USA; ⁴ Cell and Molecular Biology Group, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104, USA; ⁵ Morphochem, Inc., Monmouth Junction, New Jersey 08852, USA

Genome sequencing has been completed for multiple organisms, and pilot proteomic analyses reported for yeast and higher eukaryotes. This work has emphasized the facts that proteins are frequently engaged in multiple interactions, and that governance of protein interaction specificity is a primary means of regulating biological systems. In particular, the ability to deconvolute complex protein interaction networks to identify which interactions govern specific signaling pathways requires the generation of biological tools that allow the distinction of critical from noncritical interactions. We report the application of an enhanced Dual Bait two-hybrid system to allow detection and manipulation of highly specific protein-protein interactions. We summarize the use of this system to detect proteins and peptides that target well-defined specific motifs in larger protein structures, to facilitate rapid identification of specific interactors from a pool of putative interacting proteins obtained in a library screen, and to score specific drug-mediated disruption of protein-protein interaction.

[Supplemental material is available online at http://www.genome.org. The following individuals kindly provided reagents, samples, or unpublished information as indicated in the paper: A. Taliana, M. Russell, M. Berman, and R. Finley.]