| Chapter title |
Detecting Interactions of Membrane Proteins: The Split-Ubiquitin System
|
|---|---|
| Chapter number | 4 |
| Book title |
Two-Hybrid Systems
|
| Published in |
Methods in molecular biology, January 2018
|
| DOI | 10.1007/978-1-4939-7871-7_4 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-7870-0, 978-1-4939-7871-7
|
| Authors |
Lisa Yasmin Asseck, Christopher Grefen, Asseck, Lisa Yasmin, Grefen, Christopher |
| Abstract |
The in vivo analysis of protein-protein interactions (PPIs) is a critical factor for gaining insights into cellular mechanisms and their biological functions. To that end, a constantly growing number of genetic tools has been established, some of which are using baker's yeast (Saccharomyces cerevisiae) as a model organism. Here, we provide a detailed protocol for the yeast mating-based split-ubiquitin system (mbSUS) to study binary interactions among or with full-length membrane proteins in their native subcellular environment. The system is based on the reassembly of two autonomously non-functional ubiquitin moieties attached to proteins of interest (POIs) into a native-like molecule followed by the release of a transcription factor. Upon its nuclear import, the activation of reporter gene expression gives a visual output via growth on interaction-selective media. Additionally, we apply a modification of the classical split-ubiquitin technique called CytoSUS that detects interactions of non-membrane/soluble proteins in their full-length form via translational fusion of an ER membrane anchor. |
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Mendeley readers
Geographical breakdown
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|---|---|---|
| Unknown | 15 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
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| Professor > Associate Professor | 1 | 7% |
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