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

The Ras Gene02:38

The Ras Gene

7.4K
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...
7.4K
Small GTPases - Ras and Rho01:24

Small GTPases - Ras and Rho

5.6K
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:
5.6K
MAPK Signaling Cascades01:07

MAPK Signaling Cascades

9.0K
Mitogen-activated protein kinase, or MAPK pathway, activates three sequential kinases to regulate cellular responses such as proliferation, differentiation, survival, and apoptosis. The canonical MAPK pathway starts with a mitogen or growth factor binding to an RTK. The activated RTKs stimulate Ras, which recruits Raf or MAP3 Kinase (MAPKKK), the first kinase of the MAPK signaling cascade. Raf further phosphorylates and activates MEK or MAP2 Kinases (MAPKK), which in turn phosphorylates MAP...
9.0K
Rab Proteins01:14

Rab Proteins

5.3K
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...
5.3K
Abnormal Proliferation02:23

Abnormal Proliferation

5.4K
Under normal conditions, most adult cells remain in a non-proliferative state unless stimulated by internal or external factors to replace lost cells. Abnormal cell proliferation is a condition in which the cell's growth exceeds and is uncoordinated with normal cells. In such situations, cell division persists in the same excessive manner even after cessation of the stimuli, leading to persistent tumors. The tumor arises from the damaged cells that replicate to pass the damage to the...
5.4K
Cancer-Critical Genes I: Proto-oncogenes01:33

Cancer-Critical Genes I: Proto-oncogenes

11.7K
Genes usually encode proteins necessary for the proper functioning of a healthy cell. Mutations can often cause changes to the gene expression pattern, thereby altering the phenotype.
When the function of certain critical genes, especially those involved in cell cycle regulation and cell growth signaling cascades, gets disrupted, it upsets the cell cycle progression. Such cells with unchecked cell cycles start proliferating uncontrollably and eventually develop into tumors.
Such genes that act...
11.7K

You might also read

Related Articles

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

Sort by
Same author

Methods for the In Vitro Selection of Protein and Peptide Libraries Using mRNA Display.

Methods in molecular biology (Clifton, N.J.)·2026
Same author

Post-Selection Methods for Analyzing mRNA Display Selections and Optimization of Hits.

Methods in molecular biology (Clifton, N.J.)·2026
Same author

Integrating Diffusion and Liquid AI Models for Predicting Peptide Affinity from mRNA Display Selections.

bioRxiv : the preprint server for biology·2026
Same author

Prognostic Value of the CONUT Score in Predicting All-Cause Mortality in Hospitalized Internal Medicine Patients: A Retrospective Cohort Study.

Journal of clinical medicine·2026
Same author

When algorithmic managers fail to fulfill their promises: The role of anthropomorphism in shaping justice perceptions.

PloS one·2026
Same author

Cryo-EM of autoantibody-bound NMDA receptors reveals antigenic hotspots in an active immunization model of anti-NMDAR encephalitis.

Science advances·2026

Related Experiment Video

Updated: Mar 11, 2026

Characterize Disease-related Mutants of RAF Family Kinases by Using a Set of Practical and Feasible Methods
07:49

Characterize Disease-related Mutants of RAF Family Kinases by Using a Set of Practical and Feasible Methods

Published on: July 17, 2019

6.6K

RasIns: Genetically Encoded Intrabodies of Activated Ras Proteins.

Mehmet Cetin1, William E Evenson2, Garrett G Gross1

  • 1Department of Molecular and Computational Biology, University of Southern California, Los Angeles, CA 90089, USA.

Journal of Molecular Biology
|November 21, 2016
PubMed
Summary

Researchers developed novel antibody-like intrabodies targeting active, GTP-bound K-Ras and H-Ras. These intrabodies show high specificity and selectivity for oncogenic Ras mutants, offering potential tools for cancer research.

Keywords:
E10FnIIIRasfibronectinintrabodymRNA display

More Related Videos

Fully Processed Recombinant KRAS4b: Isolating and Characterizing the Farnesylated and Methylated Protein
07:08

Fully Processed Recombinant KRAS4b: Isolating and Characterizing the Farnesylated and Methylated Protein

Published on: January 16, 2020

6.2K
Author Spotlight: Integrating BRET-Based Assays and Rare Mutation Analysis to Decipher RAF Kinase Regulation in Live Cells
06:44

Author Spotlight: Integrating BRET-Based Assays and Rare Mutation Analysis to Decipher RAF Kinase Regulation in Live Cells

Published on: March 1, 2024

1.8K

Related Experiment Videos

Last Updated: Mar 11, 2026

Characterize Disease-related Mutants of RAF Family Kinases by Using a Set of Practical and Feasible Methods
07:49

Characterize Disease-related Mutants of RAF Family Kinases by Using a Set of Practical and Feasible Methods

Published on: July 17, 2019

6.6K
Fully Processed Recombinant KRAS4b: Isolating and Characterizing the Farnesylated and Methylated Protein
07:08

Fully Processed Recombinant KRAS4b: Isolating and Characterizing the Farnesylated and Methylated Protein

Published on: January 16, 2020

6.2K
Author Spotlight: Integrating BRET-Based Assays and Rare Mutation Analysis to Decipher RAF Kinase Regulation in Live Cells
06:44

Author Spotlight: Integrating BRET-Based Assays and Rare Mutation Analysis to Decipher RAF Kinase Regulation in Live Cells

Published on: March 1, 2024

1.8K

Area of Science:

  • Oncology
  • Molecular Biology
  • Biotechnology

Background:

  • K-Ras and H-Ras are frequently mutated in human cancers, driving tumor growth and poor prognoses.
  • Targeting oncogenic Ras proteins is a significant challenge in cancer therapy.

Purpose of the Study:

  • To design and develop genetically encoded antibody-like ligands (intrabodies) that specifically recognize active, GTP-bound K-Ras and H-Ras.
  • To create tools for monitoring and potentially modulating Ras-mediated signaling pathways.

Main Methods:

  • Utilized mRNA display for primary selection of intrabodies against Ras.
  • Employed affinity maturation to enhance intrabody binding affinity and selectivity.
  • Fused intrabodies with fluorescent proteins for cellular localization studies.

Main Results:

  • Developed RasIn1, an intrabody with micromolar affinity (KD = 2.1μM) for H-Ras(G12V)-GTP, demonstrating high state selectivity and specificity for K- and H-Ras.
  • Generated RasIn2 through affinity maturation, achieving nanomolar affinity (KD = 120nM) while maintaining selectivity.
  • Observed colocalization of both intrabodies with Ras proteins and their mutants within cells.
  • Demonstrated mutant-selective recognition of G12V Ras mutants over wild-type Ras.

Conclusions:

  • Engineered intrabodies (RasIn1 and RasIn2) effectively bind active, GTP-bound K- and H-Ras, including oncogenic mutants.
  • These intrabodies exhibit high specificity and selectivity, with potential for mutant-specific targeting.
  • The developed intrabodies serve as promising tools for investigating and potentially interfering with Ras signaling in cancer.