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Related Concept Videos

Small GTPases - Ras and Rho01:24

Small GTPases - Ras and Rho

Ras and Rho are small monomeric GTPases that act downstream of receptor tyrosine kinase (RTK) and regulate various cellular processes. These GTPases switch between active and inactive states by binding to guanine nucleotides.
Three regulatory proteins control their activity:
GTPases and their Regulation02:14

GTPases and their Regulation

Guanine nucleotide-binding proteins (G-proteins), also known as GTPases, are a superfamily of proteins that regulate many cellular processes, such as cell signaling, vesicular transport, and the regulation of cell shape and motility. Mutation or dysfunction of these proteins can lead to disease. There are around 40,000 known G-proteins that can broadly be classified into two groups ‒  small G-proteins consisting of a single domain and large multi-domain G-proteins.
Large G-proteins, also known...
GTPases and their Regulation02:14

GTPases and their Regulation

Guanine nucleotide-binding proteins (G-proteins), also known as GTPases, are a superfamily of proteins that regulate many cellular processes, such as cell signaling, vesicular transport, and the regulation of cell shape and motility. Mutation or dysfunction of these proteins can lead to disease. There are around 40,000 known G-proteins that can broadly be classified into two groups ‒  small G-proteins consisting of a single domain and large multi-domain G-proteins.
Large G-proteins, also known...
Activation and Inactivation of G Proteins01:22

Activation and Inactivation of G Proteins

Heterotrimeric G proteins are guanine nucleotide-binding proteins. As the name suggests, heterotrimeric G proteins are composed of three subunits: alpha, beta, and gamma. They remain GDP-bound or GTP-bound inside the cells and switch between inactive/active states. The Gα subunit possesses the nucleotide-binding pocket that binds guanine nucleotides and switches between GDP or GTP-bound states. In contrast, the Gꞵ and Gγ subunits are always bound together with high affinity and are together...
Rab Cascades01:25

Rab Cascades

Rab GTPases act in a regulated cascade during membrane fusion, helping the lipid bilayers mix. The Rab family of proteins are active when bound to GTP, and inactive when bound to GDP. Hence, they act as guanine nucleotide-dependent molecular switches. Rab-GTP recognizes and binds to long or short-range tethering proteins to capture the target vesicle. These tethers coordinate with SNAREs on the vesicle and the target membrane to assemble the trans SNARE complex that locks the mixing bilayers.
Rab Proteins01:14

Rab Proteins

Rab proteins constitute the largest family of monomeric GTPases, of which 70 members are present in humans. Rab proteins and their effectors regulate consecutive stages of vesicle transport such as vesicle transport, docking, and fusion to the correct recipient membrane.
Rab proteins switch between a cytosolic, GDP-bound inactive state and a membrane-anchored, GTP-bound active state. By themselves, Rabs show slow rates of GDP/GTP exchange and GTP hydrolysis. Thus, Rab proteins are considered...

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Updated: Jun 17, 2026

Detection of Small GTPase Prenylation and GTP Binding Using Membrane Fractionation and GTPase-linked Immunosorbent Assay
13:51

Detection of Small GTPase Prenylation and GTP Binding Using Membrane Fractionation and GTPase-linked Immunosorbent Assay

Published on: November 11, 2018

Glutamine Tautomerization Drives RhoGAP-Aided GTP Hydrolysis in Small Rho GTPases.

Angela Parise1, Riccardo Rozza1, Karolina Mitusinska1

  • 1CNR - Istituto Officina dei Materiali (IOM) c/o, SISSAvia Bonomea 265, Trieste34136, Italy.

Journal of the American Chemical Society
|June 15, 2026
PubMed
Summary
This summary is machine-generated.

This study reveals the molecular mechanism of GTP hydrolysis in Rho GTPases, crucial for cell motility. A specific glutamine tautomerization within the RhoGAP-RhoA complex facilitates GTP breakdown and regulates the molecular switch.

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Last Updated: Jun 17, 2026

Detection of Small GTPase Prenylation and GTP Binding Using Membrane Fractionation and GTPase-linked Immunosorbent Assay
13:51

Detection of Small GTPase Prenylation and GTP Binding Using Membrane Fractionation and GTPase-linked Immunosorbent Assay

Published on: November 11, 2018

RhoC GTPase Activation Assay
09:58

RhoC GTPase Activation Assay

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Affinity Precipitation of Active Rho-GEFs Using a GST-tagged Mutant Rho Protein (GST-RhoA(G17A)) from Epithelial Cell Lysates
11:28

Affinity Precipitation of Active Rho-GEFs Using a GST-tagged Mutant Rho Protein (GST-RhoA(G17A)) from Epithelial Cell Lysates

Published on: March 31, 2012

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Background:

  • Rho GTPases act as molecular switches regulating cell motility.
  • GTP hydrolysis by Rho GTPases is essential for their function.
  • The precise catalytic mechanism of Rho GTPases remains incompletely understood.

Purpose of the Study:

  • To elucidate the detailed molecular mechanism of GTP hydrolysis in the RhoGAP-RhoA complex.
  • To understand the role of glutamine in the catalytic process.

Main Methods:

  • Classical and hybrid quantum-classical molecular dynamics simulations.
  • Analysis of the RhoGAP-RhoA complex structure and dynamics.

Main Results:

  • GTP hydrolysis proceeds via a dissociative nucleophilic substitution mechanism.
  • An amide to imide tautomerization of Gln63 is critical for proton transfer to the phosphate group.
  • Gln63 tautomerization facilitates solvent access, enabling restoration of the active site.

Conclusions:

  • The study resolves the mechanism of GTP hydrolysis in Rho GTPases.
  • The identified mechanism involving Gln63 tautomerization may be conserved across the Rho/Rho GAP family.
  • This finding provides insights into the regulation of cytoskeletal dynamics and cell motility.