title: Genetic screen to dissect protein-protein interactions: ribonuclease inhibitor-ribonuclease A as a model system.
abstract: Protein-protein interactions are critical for the function of biological systems. Here, we describe a means to dissect a protein-protein interaction. Our method is based on the in vivo interaction between a target protein and the peptide epitopes derived from its partner. This interaction is detected by using hybrid proteins in which the target protein and peptide epitopes are fused to the DNA-binding domain of the lambda repressor protein. An interaction prevents the transcription of a reporter gene. The efficacy of this approach is demonstrated with the ribonuclease inhibitor protein and ribonuclease A, which form a complex with an equilibrium dissociation constant in the femtomolar range. Our method can enable the identification of residues important in a designated protein-protein interaction and the development of antagonists for that interaction.
authors.affiliation: Department of Biochemistry, University of Wisconsin-Madison, 433 Babcock Drive, Madison, WI 53706-1544, USA.
authors.forename: Sang-Hyun
authors.initials: SH
authors.lastname: Park
authors.forename: Ronald T
authors.initials: RT
authors.lastname: Raines
mesh_terms: D004791:Enzyme Inhibitors / Q000378:metabolism*; D000939:Epitopes; D017930:Genes, Reporter; D010957:Plasmids; D011485:Protein Binding; D012259:Ribonuclease, Pancreatic / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism*
chemical_list: D004791:Enzyme Inhibitors; D000939:Epitopes; D012259:Ribonuclease, Pancreatic