Analysis of activated GAPs and GEFs in cell lysates

Methods Enzymol. 2006;406:425-37. doi: 10.1016/S0076-6879(06)06031-9.

Abstract

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CHO Cells
  • Cricetinae
  • GTPase-Activating Proteins / metabolism*
  • Glutathione Transferase / genetics
  • Guanine Nucleotide Exchange Factors / metabolism*
  • Humans
  • Mice
  • NIH 3T3 Cells
  • Recombinant Fusion Proteins / metabolism
  • rho GTP-Binding Proteins / genetics
  • rho GTP-Binding Proteins / isolation & purification

Substances

  • GTPase-Activating Proteins
  • Guanine Nucleotide Exchange Factors
  • Recombinant Fusion Proteins
  • Glutathione Transferase
  • rho GTP-Binding Proteins