Rapid parallel flow cytometry assays of active GTPases using effector beads

Anal Biochem. 2013 Nov 15;442(2):149-57. doi: 10.1016/j.ab.2013.07.039. Epub 2013 Aug 6.


We describe a rapid assay for measuring the cellular activity of small guanine triphosphatases (GTPases) in response to a specific stimulus. Effector-functionalized beads are used to quantify in parallel multiple GTP-bound GTPases in the same cell lysate by flow cytometry. In a biologically relevant example, five different Ras family GTPases are shown for the first time to be involved in a concerted signaling cascade downstream of receptor ligation by Sin Nombre hantavirus.

Keywords: Cdc42; Hantavirus; Integrins; Rab GTPases; Rac; Rho.

Publication types

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

MeSH terms

  • Animals
  • Chlorocebus aethiops
  • Enzyme Activation
  • Enzyme Assays / methods*
  • Flow Cytometry / methods*
  • GTP Phosphohydrolases / metabolism*
  • HeLa Cells
  • Humans
  • Microspheres*
  • Single-Cell Analysis
  • Time Factors
  • Vero Cells


  • GTP Phosphohydrolases