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 temporal activation of GEFs and GAPs in response to upstream signals.
García-Mata
Rafael
R
Department of Cell and Developmental Biology, Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, USA.
Wennerberg
Krister
K
Arthur
William T
WT
Noren
Nicole K
NK
Ellerbroek
Shawn M
SM
Burridge
Keith
K
D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D020690:GTPase-Activating Proteins / Q000378:metabolism*; D005982:Glutathione Transferase / Q000235:genetics; D020662:Guanine Nucleotide Exchange Factors / Q000378:metabolism*; D006801:Humans; D051379:Mice; D041681:NIH 3T3 Cells; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D020741:rho GTP-Binding Proteins / Q000235:genetics / Q000302:isolation & purification
D020690:GTPase-Activating Proteins; D020662:Guanine Nucleotide Exchange Factors; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase; D020741:rho GTP-Binding Proteins