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A fluorescent two-hybrid assay for direct visualization of protein interactions in living cells.

Zolghadr, Kourosh and Mortusewicz, Oliver and Rothbauer, Ulrich and Kleinhans, Regina and Goehler, Heike and Wanker, Erich E. and Cardoso, M Cristina and Leonhardt, Heinrich (2008):
A fluorescent two-hybrid assay for direct visualization of protein interactions in living cells.
7, In: Molecular & cellular proteomics : MCP, (11), pp. 2279-87, ISSN 1535-9484, [Online-Edition: http://www.cardoso-lab.org/publications/Zolghadr_2008.pdf],
[Article]

Abstract

Genetic high throughput screens have yielded large sets of potential protein-protein interactions now to be verified and further investigated. Here we present a simple assay to directly visualize protein-protein interactions in single living cells. Using a modified lac repressor system, we tethered a fluorescent bait at a chromosomal lac operator array and assayed for co-localization of fluorescent prey fusion proteins. With this fluorescent two-hybrid assay we successfully investigated the interaction of proteins from different subcellular compartments including nucleus, cytoplasm, and mitochondria. In combination with an S phase marker we also studied the cell cycle dependence of protein-protein interactions. These results indicate that the fluorescent two-hybrid assay is a powerful tool to investigate protein-protein interactions within their cellular environment and to monitor the response to external stimuli in real time.

Item Type: Article
Erschienen: 2008
Creators: Zolghadr, Kourosh and Mortusewicz, Oliver and Rothbauer, Ulrich and Kleinhans, Regina and Goehler, Heike and Wanker, Erich E. and Cardoso, M Cristina and Leonhardt, Heinrich
Title: A fluorescent two-hybrid assay for direct visualization of protein interactions in living cells.
Language: English
Abstract:

Genetic high throughput screens have yielded large sets of potential protein-protein interactions now to be verified and further investigated. Here we present a simple assay to directly visualize protein-protein interactions in single living cells. Using a modified lac repressor system, we tethered a fluorescent bait at a chromosomal lac operator array and assayed for co-localization of fluorescent prey fusion proteins. With this fluorescent two-hybrid assay we successfully investigated the interaction of proteins from different subcellular compartments including nucleus, cytoplasm, and mitochondria. In combination with an S phase marker we also studied the cell cycle dependence of protein-protein interactions. These results indicate that the fluorescent two-hybrid assay is a powerful tool to investigate protein-protein interactions within their cellular environment and to monitor the response to external stimuli in real time.

Journal or Publication Title: Molecular & cellular proteomics : MCP
Volume: 7
Number: 11
Divisions: 10 Department of Biology > Cell Biology and Epigenetics
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10 Department of Biology
Date Deposited: 06 Mar 2010 16:06
Official URL: http://www.cardoso-lab.org/publications/Zolghadr_2008.pdf
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