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

7.5K
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...
7.5K
The Ras Gene02:38

The Ras Gene

2.5K
2.5K
Small GTPases - Ras and Rho01:24

Small GTPases - Ras and Rho

5.7K
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:
5.7K

You might also read

Related Articles

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

Sort by
Same author

Use of tandem-affinity chromatography to incorporate nucleotide loading of small GTPases during their purification.

Protein science : a publication of the Protein Society·2026
Same author

The scientific legacy of Martin Karplus from the perspective of his collaborators.

Biophysical journal·2026
Same author

The Neomorphic Chemistry of KRAS Autophosphorylation Is Dynamically Regulated by Active Site Conformation.

ACS chemical biology·2026
Same author

An evolutionarily conserved salt bridge stabilizes the active site for GTP hydrolysis in Rho GTPases.

The Journal of biological chemistry·2026
Same author

Ras/Raf dimerization model for activation of Raf kinase.

Current opinion in structural biology·2025
Same author

Mass spectrometry methods and mathematical PK/PD model for decision tree-guided covalent drug development.

Nature communications·2025

Related Experiment Video

Updated: Mar 26, 2026

PCR Mutagenesis, Cloning, Expression, Fast Protein Purification Protocols and Crystallization of the Wild Type and Mutant Forms of Tryptophan Synthase
09:31

PCR Mutagenesis, Cloning, Expression, Fast Protein Purification Protocols and Crystallization of the Wild Type and Mutant Forms of Tryptophan Synthase

Published on: September 26, 2020

4.9K

Expression, purification, crystallization and X-ray data collection for RAS and its mutants.

Christian W Johnson1, Greg Buhrman2, Pamela Y Ting3

  • 1Department of Chemistry and Chemical Biology, Northeastern University, Boston, MA 02115, USA.

Data in Brief
|February 12, 2016
PubMed
Summary

This study details the crystal structures of human H-RAS protein mutants, Y137E and Y137F. These structures provide insights into the effects of tyrosine phosphorylation on RAS signaling pathways.

Keywords:
Nucleotide exchangeProtein purificationRAS GTPaseX-ray crystal structures

More Related Videos

Combining X-Ray Crystallography with Small Angle X-Ray Scattering to Model Unstructured Regions of Nsa1 from S. Cerevisiae
09:15

Combining X-Ray Crystallography with Small Angle X-Ray Scattering to Model Unstructured Regions of Nsa1 from S. Cerevisiae

Published on: January 10, 2018

10.4K
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

Related Experiment Videos

Last Updated: Mar 26, 2026

PCR Mutagenesis, Cloning, Expression, Fast Protein Purification Protocols and Crystallization of the Wild Type and Mutant Forms of Tryptophan Synthase
09:31

PCR Mutagenesis, Cloning, Expression, Fast Protein Purification Protocols and Crystallization of the Wild Type and Mutant Forms of Tryptophan Synthase

Published on: September 26, 2020

4.9K
Combining X-Ray Crystallography with Small Angle X-Ray Scattering to Model Unstructured Regions of Nsa1 from S. Cerevisiae
09:15

Combining X-Ray Crystallography with Small Angle X-Ray Scattering to Model Unstructured Regions of Nsa1 from S. Cerevisiae

Published on: January 10, 2018

10.4K
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

Area of Science:

  • Structural Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Human H-RAS is a key signaling protein involved in cell growth and differentiation.
  • Tyrosine phosphorylation of H-RAS at Y137 by ABL kinases influences effector binding and downstream signaling.
  • Understanding the structural basis of Y137 phosphorylation is crucial for deciphering RAS pathway regulation.

Purpose of the Study:

  • To present detailed crystal structure data for human H-RAS Y137E and Y137F mutants.
  • To provide a structural basis for the effects of Y137 phosphorylation on H-RAS function.
  • To facilitate further research into RAS-mediated signaling.

Main Methods:

  • Expression and purification of untagged human H-RAS and its Y137E/Y137F mutants.
  • In vitro guanine nucleotide exchange assays.
  • Protein crystallization, X-ray data collection, and structure refinement.
  • Deposition of structures in the Protein Data Bank (PDB codes 4XVQ and 4XVR).

Main Results:

  • Determination of the crystal structures for H-RAS(Y137E) and H-RAS(Y137F) mutants.
  • The Y137E mutant mimics the phosphorylated state, while Y137F lacks the phosphorylation site.
  • Detailed structural information on the local environment of Y137 in H-RAS.

Conclusions:

  • The provided crystal structures offer a detailed view of H-RAS with modifications at Y137.
  • These structures serve as a valuable resource for understanding allosteric regulation of RAS effector binding.
  • Further investigation into the functional consequences of Y137 phosphorylation can be informed by these structural data.