Yeast surface two-hybrid for quantitative in vivo detection of protein-protein interactions via the secretory pathway.

ABSTRACT: A quantitative in vivo method for detecting protein-protein interactions will enhance our understanding of protein interaction networks and facilitate affinity maturation as well as designing new interaction pairs. We have developed a novel platform, dubbed "yeast surface two-hybrid (YS2H)," to enable a quantitative measurement of pairwise protein interactions via the secretory pathway by expressing one protein (bait) anchored to the cell wall and the other (prey) in soluble form. In YS2H, the prey is released either outside of the cells or remains on the cell surface by virtue of its binding to the bait. The strength of their interaction is measured by antibody binding to the epitope tag appended to the prey or direct readout of split green fluorescence protein (GFP) complementation. When

SUBMITTER: Hu X 

PROVIDER: S-EPMC2713516 | biostudies-literature | 2009 Jun

REPOSITORIES: biostudies-literature