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

Ligand Binding Sites02:40

Ligand Binding Sites

Proteins are dynamic macromolecules that carry out a wide variety of essential processes; however, the activities of most proteins depend on their interactions with other molecules or ions, known as ligands.
Protein-ligand interactions are quite specific; even though numerous potential ligands surround a cellular protein at any given time, only a particular ligand can bind to that protein. Moreover, a ligand binds only to a dedicated area on the surface of the protein, known as the...
Activation of Integrins01:15

Activation of Integrins

Integrins bind ligands and transmit information from outside the cell to inside or vice-versa through an "outside-in signaling" or "inside-out signaling."
In "outside-in signaling," external factors in the extracellular space bind to exposed ligand binding sites on integrins. This causes the inactive protein to undergo a conformational change to become active. Integrins are often clustered on the cell membrane. Repetitive and regularly spaced ligand binding events provide an effective stimulus.
Immunoglobulin-like Cell Adhesion Molecules01:31

Immunoglobulin-like Cell Adhesion Molecules

Immunoglobulin-like cell adhesion molecules or Ig-CAMs are a versatile group of cell surface glycoproteins belonging to the immunoglobulin protein superfamily. Ig-CAMs possess the characteristic immunoglobulin protein domains and other domains such as the fibronectin type III domain. The Ig domains are glycosylated to varying degrees in different Ig-CAMs.
Ig-CAMs exhibit either homophilic binding (to other Ig-CAMs) or heterophilic binding (to other ligands such as integrins). While most Ig-CAMs...
Conserved Binding Sites01:49

Conserved Binding Sites

Many proteins’ biological role depends on their interactions with their ligands, small molecules that bind to specific locations on the protein known as ligand-binding sites. Ligand-binding sites are often conserved among homologous proteins as these sites are critical for protein function.
Binding sites are often located in large pockets, and if their location on a protein’s surface is unknown, it can be predicted using various approaches. The energetic method computationally analyses the...

You might also read

Related Articles

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

Sort by
Same author

Detecting Uncoded Self-Harm in Veterans' Electronic Health Records Using Positive and Unlabeled Learning: Retrospective Cohort Study.

Journal of medical Internet research·2026
Same author

Detecting Uncoded Self-Harm in Veterans' Electronic Health Records Using Positive and Unlabeled Learning: Retrospective Observational Study.

Journal of medical Internet research·2026
Same author

The Common Fund Data Ecosystem (CFDE).

bioRxiv : the preprint server for biology·2026
Same author

KG2ML: integrating knowledge graphs and positive unlabeled learning for identifying disease-associated genes.

Frontiers in bioinformatics·2026
Same author

Trends and Predictors of Emergency Department Outcomes in Atrial Fibrillation: A Statewide Analysis from North Carolina-2016 to 2023.

Journal of the American College of Emergency Physicians open·2025
Same author

Patient-level predictors of unspecified head injury coding in traumatic brain injury-related population-based emergency department visits.

Injury prevention : journal of the International Society for Child and Adolescent Injury Prevention·2025

Related Experiment Video

Updated: Jun 20, 2026

A Flow Cytometry-Based High-Throughput Technique for Screening Integrin-Inhibitory Drugs
04:15

A Flow Cytometry-Based High-Throughput Technique for Screening Integrin-Inhibitory Drugs

Published on: February 2, 2024

Conformational mAb as a tool for integrin ligand discovery.

Ben H Njus1, Alexandre Chigaev, Anna Waller

  • 1Department of Chemistry, University of New Mexico Health Sciences Center, Albuquerque, New Mexico 87131, USA.

Assay and Drug Development Technologies
|September 17, 2009
PubMed
Summary

We developed a novel method using an anti-beta(1)-integrin antibody (HUTS-21) to measure the binding affinity of small molecule integrin ligands, aiding in the discovery of new therapeutics.

More Related Videos

Visualizing the Conformational Dynamics of Membrane Receptors Using Single-Molecule FRET
10:59

Visualizing the Conformational Dynamics of Membrane Receptors Using Single-Molecule FRET

Published on: August 17, 2022

Related Experiment Videos

Last Updated: Jun 20, 2026

A Flow Cytometry-Based High-Throughput Technique for Screening Integrin-Inhibitory Drugs
04:15

A Flow Cytometry-Based High-Throughput Technique for Screening Integrin-Inhibitory Drugs

Published on: February 2, 2024

Visualizing the Conformational Dynamics of Membrane Receptors Using Single-Molecule FRET
10:59

Visualizing the Conformational Dynamics of Membrane Receptors Using Single-Molecule FRET

Published on: August 17, 2022

Area of Science:

  • Biochemistry
  • Immunology
  • Pharmacology

Background:

  • alpha(4)beta(1)-Integrin (VLA-4) mediates leukocyte adhesion and extravasation, crucial in inflammatory diseases.
  • Small molecules blocking VLA-4-ligand interactions are potential therapeutics.
  • Existing methods for ligand binding affinity assessment can be limiting.

Purpose of the Study:

  • To establish a novel method for quantifying the binding affinity of small molecule VLA-4 ligands.
  • To validate this method using known and novel integrin ligands.
  • To explore its potential for high-throughput screening.

Main Methods:

  • Utilized a conformationally sensitive antibody (HUTS-21) and a fluorescent ligand (LDV-FITC) in flow cytometry.
  • Assessed HUTS-21 epitope exposure induced by small molecule binding.
  • Determined EC(50) values for HUTS-21 binding and correlated them with K(i) values from competition assays.

Main Results:

  • Small molecule binding to VLA-4 induced HUTS-21 epitope exposure.
  • HUTS-21 binding EC(50) values correlated well with K(i) values for tested ligands.
  • A docking model suggested a common binding mode for small molecule VLA-4 ligands.

Conclusions:

  • HUTS-21 binding serves as a reliable indicator of ligand-binding site occupancy.
  • This novel approach enables affinity determination for unlabeled integrin ligands.
  • The method is adaptable for high-throughput screening to identify novel integrin ligands.