Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Small GTPases - Ras and Rho01:24

Small GTPases - Ras and Rho

3.9K
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:
3.9K
The Ras Gene02:38

The Ras Gene

6.1K
The Ras-gene-encoded proteins are regulators of signaling pathways controlling cell proliferation, differentiation, or cell survival. The Ras-gene family in humans constitutes three primary members—the HRas, NRas, and KRas. These genes code for four functionally distinct yet closely related proteins—the HRas, NRas, KRas4A, and KRas4B. The involvement of mutant Ras genes in human cancer was first discovered in 1982 and is among the most common causes of human tumorigenesis.
Ras is a...
6.1K
Rab Cascades01:25

Rab Cascades

2.6K
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.
2.6K
Rab Proteins01:14

Rab Proteins

3.8K
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...
3.8K
Activation and Inactivation of G Proteins01:22

Activation and Inactivation of G Proteins

6.6K
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...
6.6K
GPCRs Regulate Adenylyl Cylase Activity01:09

GPCRs Regulate Adenylyl Cylase Activity

5.2K
Some GPCRs transmit signals through adenylyl cyclase (AC), a transmembrane enzyme. AC helps synthesize second messenger cyclic adenosine monophosphate (cAMP). AC catalyzes cyclization reaction and converts ATP to cAMP by releasing a pyrophosphate. The pyrophosphate is further hydrolyzed to phosphate by the enzyme pyrophosphatase, which drives cAMP synthesis to completion. However, cAMP is rapidly degraded to 5′ AMP by the enzymes phosphodiesterase (PDE), preventing overstimulation of...
5.2K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Two-step mechanism of Bruton's tyrosine kinase membrane recruitment and activation.

Proceedings of the National Academy of Sciences of the United States of America·2026
Same author

Author Correction: Mitochondrial complex II in intestinal epithelial cells regulates T cell-mediated immunopathology.

Nature immunology·2026
Same author

Two-Step Mechanism of Bruton's Tyrosine Kinase Membrane Recruitment and Activation.

bioRxiv : the preprint server for biology·2025
Same author

Clinically observed <i>RASA1</i> missense mutants exhibit diverse RasGAP protein behaviors.

bioRxiv : the preprint server for biology·2025
Same author

Nickel Binding to the c-Src SH3 Domain Facilitates Crystallization.

Protein and peptide letters·2025
Same author

Prevalence of Helicobacter pylori, Salmonella typhi, Plasmodium falciparum, and Toxoplasma gondii infections and levels of liver function markers among Hepatitis B Virus infected Ghanaians: A cross-sectional study in the Greater Accra Region.

PLOS global public health·2025

Related Experiment Video

Updated: May 29, 2025

Comparing the Affinity of GTPase-binding Proteins using Competition Assays
10:37

Comparing the Affinity of GTPase-binding Proteins using Competition Assays

Published on: October 8, 2015

9.1K

The C2 domain augments Ras GTPase-activating protein catalytic activity.

Maxum E Paul1, Di Chen2, Kimberly J Vish1

  • 1Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06520.

Proceedings of the National Academy of Sciences of the United States of America
|February 3, 2025
PubMed
Summary

The C2 domain of Ras GTPase-activating proteins (GAPs) is crucial for their signaling function. This domain enhances RasGAP catalytic activity by interacting with Ras, impacting vascular development.

Keywords:
GTP hydrolysisGTPase-activating proteinRASA1Ras signalingp120RasGAP

More Related Videos

RhoC GTPase Activation Assay
09:58

RhoC GTPase Activation Assay

Published on: August 22, 2010

12.4K
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

9.8K

Related Experiment Videos

Last Updated: May 29, 2025

Comparing the Affinity of GTPase-binding Proteins using Competition Assays
10:37

Comparing the Affinity of GTPase-binding Proteins using Competition Assays

Published on: October 8, 2015

9.1K
RhoC GTPase Activation Assay
09:58

RhoC GTPase Activation Assay

Published on: August 22, 2010

12.4K
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

9.8K

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cell Signaling

Background:

  • Ras GTPases are key regulators of cellular signaling pathways.
  • GTPase-activating proteins (GAPs) control Ras activity by promoting GTP hydrolysis.
  • Many Ras GAPs possess a C2 domain adjacent to their catalytic GAP domain.

Purpose of the Study:

  • To investigate the role of the C2 domain in RasGAP's catalytic activity.
  • To elucidate the structural basis for C2 domain-mediated regulation of RasGAP.
  • To determine the in vivo significance of the C2 domain in RasGAP function.

Main Methods:

  • Analysis of RasGAP C2-GAP crystal structure and AlphaFold models.
  • Sequence conservation analysis to identify key residues.
  • In vitro biochemical assays to measure catalytic activity.
  • In vivo studies using mouse models with specific mutations.

Main Results:

  • The C2 domain of RasGAP is essential for its full catalytic activity toward Ras.
  • Structural and sequence analyses reveal direct interaction between the RasGAP C2 domain and the Ras allosteric lobe.
  • A conserved surface on the C2 domain, involving residue R707, is critical for this interaction and catalytic enhancement.
  • Mutation of R707 in mice recapitulates vascular and signaling defects observed in RASA1 disruption.
  • Similar findings were observed for SynGAP, indicating a conserved mechanism.

Conclusions:

  • The C2 domain is a critical regulatory module for RasGAP family proteins, enhancing their catalytic efficiency.
  • The interaction between the C2 domain and Ras is evolutionarily conserved and essential for proper Ras signaling.
  • Understanding this mechanism provides insights into RasGAP function and associated diseases like vascular malformations.