A Genetic System Based on Split-Ubiquitin for the Analysis of Interactions between Membrane Proteins in vivo

A detection system for interactions between membrane proteins in vivo is described. The system is based on split-ubiquitin [Johnson, N. & Varshavsky, A. (1994) Proc. Natl. Acad. Sci. USA 91, 10340-10344]. Interaction between two membrane proteins is detected by proteolytic cleavage of a protein...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1998-04, Vol.95 (9), p.5187-5192
Main Authors: Stagljar, Igor, Korostensky, Chantal, Johnsson, Nils, Heesen, Stephan Te
Format: Article
Language:English
Subjects:
Amino acids
Biological Sciences
Cell growth
Cloning, Molecular - methods
Endopeptidases - metabolism
Genetic Vectors
Genetics
Herpes Simplex Virus Protein Vmw65 - metabolism
Hexosyltransferases
Journalism
Membrane proteins
Membrane Proteins - chemistry
Membranes
Plasmids
Protein Binding
Proteins
Recombinant Fusion Proteins - metabolism
Reporter genes
Saccharomyces cerevisiae
Transferases - metabolism
Ubiquitin-Specific Proteases
Ubiquitins
Ubiquitins - chemistry
Yeasts
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Summary:A detection system for interactions between membrane proteins in vivo is described. The system is based on split-ubiquitin [Johnson, N. & Varshavsky, A. (1994) Proc. Natl. Acad. Sci. USA 91, 10340-10344]. Interaction between two membrane proteins is detected by proteolytic cleavage of a protein fusion. The cleavage releases a transcription factor, which activates reporter genes in the nucleus. As a result, interaction between membrane proteins can be analyzed by the means of a colorimetric assay. We use membrane proteins of the endoplasmic reticulum as a model system. Wbp1p and Ost1p are both subunits of the oligosaccharyl transferase membrane protein complex. The Alg5 protein also localizes to the membrane of the endoplasmic reticulum, but does not interact with the oligosaccharyltransferase. Specific interactions are detected between Wbp1p and Ost1p, but not between Wbp1p and Alg5p. The new system might be useful as a genetic and biochemical tool for the analysis of interactions between membrane proteins in vivo.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.95.9.5187