Applying bimolecular fluorescence complementation to screen and purify aquaporin protein:protein complexes.

ABSTRACT: Protein:protein interactions play key functional roles in the molecular machinery of the cell. A major challenge for structural biology is to gain high-resolution structural insight into how membrane protein function is regulated by protein:protein interactions. To this end we present a method to express, detect, and purify stable membrane protein complexes that are suitable for further structural characterization. Our approach utilizes bimolecular fluorescence complementation (BiFC), whereby each protein of an interaction pair is fused to nonfluorescent fragments of yellow fluorescent protein (YFP) that combine and mature as the complex is formed. YFP thus facilitates the visualization of protein:protein interactions in vivo, stabilizes the assembled complex, and provides a fluorescent ma

SUBMITTER: Sjohamn J 

PROVIDER: S-EPMC5119558 | biostudies-literature | 2016 Dec

REPOSITORIES: biostudies-literature