title: Functional analysis of Rho GTPase activation and inhibition in a bead-based miniaturized format.
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.
authors.affiliation: Department of Biochemistry, NMI Natural and Medical Sciences Institute at the University of Tübingen, Reutlingen, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Schmohl
authors.forename: Stefanie
authors.initials: S
authors.lastname: Rimmele
authors.forename: Peter
authors.initials: P
authors.lastname: Gierschik
authors.forename: Thomas O
authors.initials: TO
authors.lastname: Joos
authors.forename: Nicole
authors.initials: N
authors.lastname: Schneiderhan-Marra
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism; D001667:Binding, Competitive; D003546:Cysteine Endopeptidases / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D008863:Microspheres; D009711:Nucleotides / Q000737:chemistry / Q000378:metabolism; D040081:Protein Array Analysis / Q000379:methods*; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D059748:Proteolysis; D020741:rho GTP-Binding Proteins / Q000819:agonists* / Q000037:antagonists & inhibitors*
chemical_list: D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D009711:Nucleotides; C116117:YopT protein, Yersinia; D003546:Cysteine Endopeptidases; D020741:rho GTP-Binding Proteins