Abstract
Extensive knowledge about protein–protein interactions is fundamental to fully understand signaling pathways and for the development of new drugs. Rho GTPases are key molecules in cellular signaling processes and their deregulation is implicated in the development of a variety of diseases such as neurofibromatosis type 2 and cancer. Here, we describe a bead-based protein–protein interaction assay for overexpressed HA-tagged Rho GTPases to study the GTPγS-dependent interaction with the regulatory protein RhoGDIα. This assay provides a useful tool for the analysis of both macromolecular and small molecule activators and inhibitors of the protein–protein interactions of Rho GTPases with their regulatory proteins in a multiplexed miniaturized format.
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Acknowledgments
This work was supported by the Deutsche Forschungsgemeinschaft (DFG), grants JO 687/2-1 and GI138/5-1. We are grateful to Prof. Yoel Kloog, Department of Neurobiochemistry, Tel Aviv University, Israel, for providing the prenyl derivatives.
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Schmohl, M., Rimmele, S., Gierschik, P., Joos, T.O., Schneiderhan-Marra, N. (2012). Functional Analysis of Rho GTPase Activation and Inhibition in a Bead-Based Miniaturized Format. In: Rivero, F. (eds) Rho GTPases. Methods in Molecular Biology, vol 827. Springer, New York, NY. https://doi.org/10.1007/978-1-61779-442-1_18
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DOI: https://doi.org/10.1007/978-1-61779-442-1_18
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