Desired alteration of protein affinities: competitive selection of protein variants using yeast signal transduction machinery.

ABSTRACT: Molecules that can control protein-protein interactions (PPIs) have recently drawn attention as new drug pipeline compounds. Here, we report a technique to screen desirable affinity-altered (affinity-enhanced and affinity-attenuated) protein variants. We previously constructed a screening system based on a target protein fused to a mutated G-protein γ subunit (Gγcyto) lacking membrane localization ability. This ability, required for signal transmission, is restored by recruiting Gγcyto into the membrane only when the target protein interacts with an artificially membrane-anchored candidate protein, thereby allowing interacting partners (Gγ recruitment system) to be searched and identified. In the present study, the Gγ recruitment system was altered by integrating the cytosolic expression o

SUBMITTER: Kaishima M 

PROVIDER: S-EPMC4171513 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature