Electrostatic forces mediate the specificity of RHO GTPase-GDI interactions

Niloufar Mosaddeghzadeh, Neda S. Kazemein Jasemi, Jisca Majolée, Si-Cai Zhang, Peter L. Hordijk, Radovan Dvorsky, Mohammad Reza Ahmadian

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5 Citations (Scopus)


Three decades of research have documented the spatiotemporal dynamics of RHO family GTPase membrane extraction regulated by guanine nucleotide dissociation inhibitors (GDIs), but the interplay of the kinetic mechanism and structural specificity of these interactions is as yet unresolved. To address this, we reconstituted the GDI-controlled spatial segregation of geranylger-anylated RHO protein RAC1 in vitro. Various biochemical and biophysical measurements provided unprecedented mechanistic details for GDI function with respect to RHO protein dynamics. We de-termined that membrane extraction of RHO GTPases by GDI occurs via a 3-step mechanism: (1) GDI non-specifically associates with the switch regions of the RHO GTPases; (2) an electrostatic switch determines the interaction specificity between the C-terminal polybasic region of RHO GTPases and two distinct negatively-charged clusters of GDI1; (3) a non-specific displacement of geranylgeranyl moiety from the membrane sequesters it into a hydrophobic cleft, effectively shielding it from the aqueous milieu. This study substantially extends the model for the mechanism of GDI-regulated RHO GTPase extraction from the membrane, and could have implications for clinical studies and drug development.
Original languageEnglish
Article number12493
JournalInternational journal of molecular sciences
Issue number22
Publication statusPublished - 1 Nov 2021


  • CDC42
  • Electrostatic steering
  • G domain
  • Geranylgeranyl
  • Guanine nucleotide dissociation inhibitors
  • Hypervariable region
  • Liposomes
  • Membrane extraction
  • Polybasic motif
  • RAC1
  • RAC2
  • RHOA

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