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

Mapping the integrin alpha V beta 3-ligand interface by photoaffinity cross-linking.

G Bitan1, L Scheibler, Z Greenberg

  • 1Division of Bone and Mineral Metabolism, Charles A. Dana and Thorndike Laboratories, Department of Medicine, Beth Israel Deaconess Medical Center, Boston, Massachusetts 02215, USA.

Biochemistry
|March 17, 1999
PubMed
Summary

Researchers developed novel photoaffinity probes to map the RGD-binding site on integrin alpha V beta 3. These probes successfully cross-linked to the beta 3 subunit, identifying a specific contact domain for ligand binding.

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

Expression ratio of the TGFβ-inducible gene MYO10 is prognostic for overall survival of squamous cell lung cancer patients and predicts chemotherapy response.

Scientific reports·2018
Same author

Guideline. Diagnosis and treatment of venous malformations. consensus document of the international union of phlebology (iup): updated-2013.

International angiology : a journal of the International Union of Angiology·2014
Same author

Diagnosis and Treatment of Venous Malformations. Consensus Document of the International Union of Phlebology (IUP): updated 2013.

International angiology : a journal of the International Union of Angiology·2014
Same author

Locomotor hyperactivity in 14-3-3ζ KO mice is associated with dopamine transporter dysfunction.

Translational psychiatry·2013
Same author

The PCAST report: impact and implications for the pharmaceutical industry.

Clinical pharmacology and therapeutics·2013
Same author

Gynecological chair syncope in elderly women: practical implications for general practitioners and gynecologists.

The International journal of risk & safety in medicine·2013

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Background:

  • Integrins are crucial cell surface adhesion receptors mediating cell-extracellular matrix interactions.
  • High-resolution structural data for integrins remain limited, hindering detailed understanding of ligand binding.
  • Previous cross-linking studies identified large binding domains but lacked precision.

Purpose of the Study:

  • To design and synthesize novel photoreactive ligands for photoaffinity scanning of the RGD-binding site on human integrin alpha V beta 3.
  • To create molecular tools for precise mapping of integrin-ligand interactions at the molecular level.

Main Methods:

  • Design of conformationally constrained, RGD-containing ligands incorporating a benzophenone moiety (Bpa) for photoactivation.
  • Introduction of molecular tags (radioactive iodine or biotin) for detection and analysis.

Related Experiment Videos

  • Photoaffinity cross-linking experiments with purified integrin alpha V beta 3.
  • Analysis of cross-linked products using SDS-PAGE, enzymatic digestion, and chemical cleavage.

    • Main Results:

    • Newly designed tagged photoreactive RGD ligands exhibited high affinity (0.5-0.7 microM) for integrin alpha V beta 3.
    • Efficient and specific cross-linking of ligands to the integrin was achieved.
    • A 100 kDa band, corresponding to the beta 3 subunit-ligand conjugate, was identified as the major cross-linking product.
    • Identification of a 20-amino acid fragment (residues 99-118) in the beta 3 chain as the ligand contact domain.

    Conclusions:

    • The developed photoaffinity ligands are effective tools for mapping integrin-ligand interfaces with high precision.
    • The study precisely localized the RGD-binding site on the beta 3 subunit of integrin alpha V beta 3.
    • This methodology advances the understanding of integrin structure-function relationships and receptor-ligand interactions.