Unit

UNIT 5.27 Split-Ubiquitin System for Identifying Protein-Protein Interactions in Membrane and Full-Length Proteins

  1. Christopher Grefen1,
  2. Sylvie Lalonde2,
  3. Petr Obrdlik3

Published Online: 1 OCT 2007

DOI: 10.1002/0471142301.ns0527s41

Current Protocols in Neuroscience

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.

Author Information

  1. 1

    Universität Tübingen, Tübingen, Germany

  2. 2

    Carnegie Institution, Stanford, California

  3. 3

    IonGate Biosciences GmbH, Frankfurt, Germany

Publication History

  1. Published Online: 1 OCT 2007
  2. Published Print: OCT 2007

Abstract

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.

Keywords:

  • Split-ubiquitin;
  • mbSUS;
  • protein-protein interaction;
  • full-length membrane protein;
  • mating;
  • systematic analysis;
  • Gateway