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 Experiment Videos

A screening method of SH2 domain ligands and blockers using a solid phase binding

W S Koh1, S Y Yoon, E K Lee

  • 1Immune Regulation Research Unit, Korea Research Institute of Bioscience and Biotechnology, Yusung, Taejon, South Korea.

Cancer Letters
|May 7, 1998
PubMed
Summary

We developed an inexpensive, rapid assay to screen for molecules that bind to the Grb2 SH2 domain. This high-throughput method is ideal for identifying potential anticancer drug candidates by analyzing signal transduction pathways.

Related Concept Videos

You might also read

Related Articles

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

Sort by
Same author

Limits on WIMP Dark Matter with NaI(Tl) Crystals in Three Years of COSINE-100 Data.

Physical review letters·2025
Same author

Combined Annual Modulation Dark Matter Search with COSINE-100 and ANAIS-112.

Physical review letters·2025
Same author

Improved Limit on Neutrinoless Double Beta Decay of ^{100}Mo from AMoRE-I.

Physical review letters·2025
Same author

Search for Boosted Dark Matter in COSINE-100.

Physical review letters·2023
Same author

Detection efficiency calibration for an array of fourteen HPGe detectors.

Applied radiation and isotopes : including data, instrumentation and methods for use in agriculture, industry and medicine·2023
Same author

Mono-PEGylates of exenatide in branched and dimeric structures can improve in vivo stability and hypoglycemic bioactivity.

Journal of biotechnology·2019

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Drug Discovery

Background:

  • SH2 domains are crucial in signal transduction pathways.
  • Grb2 SH2 domain interactions regulate cellular signaling.
  • Developing efficient screening methods for SH2 domain ligands is essential.

Purpose of the Study:

  • To develop a high-throughput screening method for SH2 domain binding ligands and blockers.
  • To characterize the binding of a phosphopeptide to the Grb2 SH2 domain.

Main Methods:

  • Utilized a 3H-labeled phosphopeptide derived from human Shc (SpYVNVK) to measure binding to Grb2 SH2 domain-glutathione S-transferase fusion proteins.
  • Optimized fusion protein coating concentration (300 ng/100 microl/well) and incubation time (8h at 4°C).
  • Determined optimal ligand peptide concentration (2 pmol/well) for reproducible binding.

Related Experiment Videos

Main Results:

  • Established optimal conditions for fusion protein coating and ligand peptide binding.
  • Demonstrated high specificity of the assay for peptide sequences.
  • Competitive inhibition studies confirmed the assay's ability to screen ligands and blockers.

Conclusions:

  • The developed method is easy, rapid, and inexpensive for SH2 domain binding assays.
  • This high-throughput assay is suitable for screening anticancer drug candidates.
  • The assay effectively screens ligands and blockers of Grb2-mediated signal transduction pathways.