Analysis of Activated GAPs and GEFs in Cell Lysates

An assay was developed that allows the precipitation of the active pools of Rho‐GEFs, Rho‐GAPs, or effectors from cell or tissue lysates. This assay can be used to identify GEFs, GAPs, and effectors involved in specific cellular pathways to determine their GTPase specificity and to monitor the tempo...

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Veröffentlicht in:	Methods in Enzymology 2006, Vol.406, p.425-437
Hauptverfasser:	García‐Mata, Rafael, Wennerberg, Krister, Arthur, William T., Noren, Nicole K., Ellerbroek, Shawn M., Burridge, Keith
Format:	Artikel
Sprache:	eng
Schlagworte:	Animals CHO Cells Cricetinae Glutathione Transferase - genetics GTPase-Activating Proteins - metabolism Guanine Nucleotide Exchange Factors - metabolism Humans Mice Molecular biology NIH 3T3 Cells Proteins Recombinant Fusion Proteins - metabolism rho GTP-Binding Proteins - genetics rho GTP-Binding Proteins - isolation & purification
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Beschreibung
Zusammenfassung:	An assay was developed that allows the precipitation of the active pools of Rho‐GEFs, Rho‐GAPs, or effectors from cell or tissue lysates. This assay can be used to identify GEFs, GAPs, and effectors involved in specific cellular pathways to determine their GTPase specificity and to monitor the temporal activation of GEFs and GAPs in response to upstream signals.
ISSN:	0076-6879 1557-7988
DOI:	10.1016/S0076-6879(06)06031-9