Protein–protein‐interactions in a multiplexed,miniaturized format a functional analysis of Rho GTPase activation and inhibition |
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| Authors: | Michael Schmohl Stefanie Rimmele Oliver Pötz Yoel Kloog Peter Gierschik Thomas O. Joos Nicole Schneiderhan‐Marra |
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| Affiliation: | 1. NMI Natural and Medical Sciences Institute at the University of Tuebingen, Reutlingen, Germany;2. Department of Neurobiochemistry, The George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv, Israel;3. Institute of Pharmacology and Toxicology, University of Ulm Medical Center, Ulm, Germany |
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| Abstract: | A miniaturized, bead‐based protein–protein‐interaction assay was developed to study the interaction of Rho GTPases with regulatory proteins. The setup, which uses only minute amounts of sample, was used to analyze small molecules that inhibit the interaction between Rho GTPases and RhoGDIα. Prenylcysteine analogues and the replacement of GDP by non‐hydrolysable GTP analogues prevented the formation of Rho GTPase‐RhoGDIα complexes in a concentration‐dependent manner. |
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| Keywords: | Automation Bead‐based assay Prenylcysteine analogues Protein arrays Rac RhoGDI |
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