title: Improved split-ubiquitin screening technique to identify surface membrane protein-protein interactions.
abstract: Yeast-based methods are still the workhorse for the detection of protein-protein interactions (PPIs) in vivo. Yeast two-hybrid (Y2H) systems, however, are limited to screening for a specific group of molecules that interact in a particular cell compartment. For this reason, the split-ubiquitin system (SUS) was developed to allow screening of cDNA libraries of full-length membrane proteins for protein-protein interactions in Saccharomyces cerevisiae. Here we demonstrate that a modification of the widely used membrane SUS involving the transmembrane (TM) domain of the yeast receptor Wsc1 increases the stringency of screening and improves the selectivity for proteins localized in the plasma membrane (PM).
authors.affiliation: Robert Koch Institute, Berlin, Germany.|Freie Universität Berlin, Berlin, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Ivanusic
authors.affiliation: Universität Osnabrück, Osnabrück, Germany.
authors.forename: Jürgen J
authors.initials: JJ
authors.lastname: Heinisch
authors.affiliation: Robert Koch Institute, Berlin, Germany.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Eschricht
authors.affiliation: Robert Koch Institute, Berlin, Germany.
authors.forename: Ulrike
authors.initials: U
authors.lastname: Laube
authors.affiliation: Robert Koch Institute, Berlin, Germany.
authors.forename: Joachim
authors.initials: J
authors.lastname: Denner
mesh_terms: D015723:Gene Library; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism*; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D012441:Saccharomyces cerevisiae; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D020798:Two-Hybrid System Techniques; D025801:Ubiquitin
chemical_list: D008565:Membrane Proteins; C119383:SLG1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D025801:Ubiquitin
keywords: protein-protein interactions; receptors; split-ubiquitin system