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Related Concept Videos

GTPases and their Regulation02:14

GTPases and their Regulation

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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.
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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.
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The bonds of adenosine triphosphate (ATP) can be broken through the addition of water, releasing one or two phosphate groups in an exergonic process called hydrolysis. This reaction liberates the energy in the bonds for use in the cell—for instance, to synthesize proteins from amino acids.
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Overview
Hydrolysis is a chemical reaction in which the addition of water breaks down a polymer into its simpler monomer units. For example, peptides break into amino acids, carbohydrates into simple sugars, and DNA into nucleotides. Enzymes often facilitate these processes.
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Conservation biology is a scientific field that focuses on the preservation of biodiversity in order to protect ecosystems while meeting the needs of the human population. Humans require properly functioning ecosystems to maintain our supply of natural resources, including food, medicines, and building materials.
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Related Experiment Video

Updated: Feb 9, 2026

Detection of Small GTPase Prenylation and GTP Binding Using Membrane Fractionation and GTPase-linked Immunosorbent Assay
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An evolutionarily conserved salt bridge stabilizes the active site for GTP hydrolysis in Rho GTPases.

Kendra Marcus1, Michael Schwabe1, Ryan Knihtila1

  • 1Department of Chemistry and Chemical Biology, Northeastern University, Boston, Massachusetts, USA.

The Journal of Biological Chemistry
|February 7, 2026
PubMed
Summary

A conserved salt bridge in Rho GTPases is crucial for stabilizing the active site and GTP hydrolysis. Disrupting this bridge in RhoA has severe effects, unlike in H-Ras, highlighting its importance for cancer drug discovery.

Keywords:
molecular dynamicsmolecular evolutionsmall GTPasestructural biologyx-ray crystallography

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Structural Biology

Background:

  • Rho GTPases regulate fundamental cellular processes like morphology, motility, and cell cycling.
  • Dysregulation of Rho GTPases is linked to various cancers, making them potential drug targets.
  • The switch II pocket is a key region for drug discovery in Ras-family GTPases.

Purpose of the Study:

  • To investigate the evolutionary trajectory and functional role of the switch II pocket in Rho GTPases.
  • To characterize the stabilizing role of a conserved salt bridge in the RhoA active site.
  • To explore the allosteric communication within Rho GTPases and its implications for drug development.

Main Methods:

  • X-ray crystallography to determine structural changes.
  • Accelerated molecular dynamics simulations (aMD) to study protein dynamics.
  • Enzymatic assays to measure GTP hydrolysis rates.

Main Results:

  • A conserved salt bridge involving R70 in RhoA stabilizes the active site for efficient GTP hydrolysis.
  • Removal of R70 in RhoA leads to catastrophic effects on active site organization and hydrolysis.
  • Removal of the analogous residue in H-Ras has more moderate effects, indicating subfamily-specific mechanisms.
  • Allosteric interactions involving residues K98 and D13 modulate activity through the salt bridge.

Conclusions:

  • The Rho GTPase salt bridge is essential for maintaining active site conformation and catalytic function.
  • Understanding the switch II pocket's functional landscape in Rho GTPases offers new avenues for cancer drug discovery.
  • Allosteric networks involving the salt bridge are critical for Rho GTPase regulation.