title: Identification of new interacting partners for atypical Rho GTPases: a SILAC-based approach.
abstract: In contrast to typical Rho GTPases the regulation of atypical Rho GTPases, such as the members of the RhoBTB subfamily, rarely depends on GEFs and/or GAPs. Instead, they are regulated at the level of their expression, by post-translational modifications, by their rate of degradation as well as through binding of diverse cell-specific interactors. Stable Isotope Labeling by Amino acids in Cell culture (SILAC) is a powerful cutting-edge mass-spectrometry-based technology allowing for protein-interaction studies in vitro with removal of false-positive identifications. In this chapter, we describe how the SILAC technology can be applied to the identification of new interacting partners for atypical - constitutively active - Rho GTPases, i.e. RhoBTB3.
authors.affiliation: Instituto de Biologia Molecular e Celular, Universidade do Porto, Porto, Portugal.
authors.forename: Laura
authors.initials: L
authors.lastname: Montani
authors.forename: Damaris
authors.initials: D
authors.lastname: Bausch-Fluck
authors.forename: Ana Filipa
authors.initials: AF
authors.lastname: Domingues
authors.forename: Bernd
authors.initials: B
authors.lastname: Wollscheid
authors.forename: João Bettencourt
authors.initials: JB
authors.lastname: Relvas
mesh_terms: D000596:Amino Acids / Q000737:chemistry* / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods; D057809:HEK293 Cells; D006801:Humans; D007553:Isotope Labeling / Q000379:methods*; D013058:Mass Spectrometry; D010957:Plasmids / Q000235:genetics; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D014162:Transfection; D020741:rho GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D000596:Amino Acids; D020741:rho GTP-Binding Proteins