UNIT 5.27 Split-Ubiquitin System for Identifying Protein-Protein Interactions in Membrane and Full-Length Proteins
Published Online: 1 OCT 2007
Copyright © 2007 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Neuroscience
How to Cite
Grefen, C., Lalonde, S. and Obrdlik, P. 2007. Split-Ubiquitin System for Identifying Protein-Protein Interactions in Membrane and Full-Length Proteins. Current Protocols in Neuroscience. 41:5.27:5.27.1–5.27.41.
- Published Online: 1 OCT 2007
- Published Print: OCT 2007
Protein-protein interactions play a fundamental role in the regulation of almost all cellular processes. Thus, the identification of interacting proteins can help to elucidate their function. The mating-based split-ubiquitin system (mbSUS) uses yeast as a test organism to identify potential interactions between full-length membrane proteins or between a full-length membrane protein and a soluble protein. The mbSUS can also be used to provide further evidence for protein-protein interactions detected with other methods and to map the interaction domains of selected proteins. The mbSUS is optimized for systematic screening approaches employing a mating-based approach, as typically used to determine protein interactions on a genomic scale. Construction of bait and prey fusions is simplified by adapting two different cloning procedures: (i) in vivo cloning in yeast, and (ii) Gateway cloning in E. coli. Protocols for small-scale interaction tests, as well as systematic approaches using sorted bait and prey arrays, are described. Curr. Protoc. Neurosci. 41:5.27.1-5.27.41. © 2007 by John Wiley & Sons, Inc.
- protein-protein interaction;
- full-length membrane protein;
- systematic analysis;