Improved split-ubiquitin screening technique to identify surface membrane protein-protein interactions
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 syste...
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2015-08-01
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| Series: | BioTechniques |
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| Online Access: | https://www.future-science.com/doi/10.2144/000114315 |
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| author | Daniel Ivanusic Jürgen J. Heinisch Magdalena Eschricht Ulrike Laube Joachim Denner |
| author_facet | Daniel Ivanusic Jürgen J. Heinisch Magdalena Eschricht Ulrike Laube Joachim Denner |
| author_sort | Daniel Ivanusic |
| collection | DOAJ |
| description | 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). |
| format | Article |
| id | doaj-art-6727c034d1ef46eea8023f028bdd5163 |
| institution | OA Journals |
| issn | 0736-6205 1940-9818 |
| language | English |
| publishDate | 2015-08-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj-art-6727c034d1ef46eea8023f028bdd51632025-08-20T02:26:06ZengTaylor & Francis GroupBioTechniques0736-62051940-98182015-08-01592637310.2144/000114315Improved split-ubiquitin screening technique to identify surface membrane protein-protein interactionsDaniel Ivanusic0Jürgen J. Heinisch1Magdalena Eschricht2Ulrike Laube3Joachim Denner41Robert Koch Institute, Berlin, Germany3Universität Osnabrück, Osnabrück, Germany1Robert Koch Institute, Berlin, Germany1Robert Koch Institute, Berlin, Germany1Robert Koch Institute, Berlin, GermanyYeast-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).https://www.future-science.com/doi/10.2144/000114315protein-protein interactionssplit-ubiquitin systemreceptors |
| spellingShingle | Daniel Ivanusic Jürgen J. Heinisch Magdalena Eschricht Ulrike Laube Joachim Denner Improved split-ubiquitin screening technique to identify surface membrane protein-protein interactions BioTechniques protein-protein interactions split-ubiquitin system receptors |
| title | Improved split-ubiquitin screening technique to identify surface membrane protein-protein interactions |
| title_full | Improved split-ubiquitin screening technique to identify surface membrane protein-protein interactions |
| title_fullStr | Improved split-ubiquitin screening technique to identify surface membrane protein-protein interactions |
| title_full_unstemmed | Improved split-ubiquitin screening technique to identify surface membrane protein-protein interactions |
| title_short | Improved split-ubiquitin screening technique to identify surface membrane protein-protein interactions |
| title_sort | improved split ubiquitin screening technique to identify surface membrane protein protein interactions |
| topic | protein-protein interactions split-ubiquitin system receptors |
| url | https://www.future-science.com/doi/10.2144/000114315 |
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