Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

The Equilibrium Binding Constant and Binding Strength

The Equilibrium Binding Constant and Binding Strength

The equilibrium binding constant (Kb) quantifies the strength of a protein-ligand interaction. Kb can be calculated as follows when the reaction is at equilibrium:

Protein-Drug Binding: Determination Methods

Protein-Drug Binding: Determination Methods

Determining protein-drug binding can be achieved through indirect and direct methods, each providing valuable insights into the interaction between proteins and drugs.
Indirect methods involve isolating the bound drug from its free form in biological samples such as blood, serum, or plasma. These techniques aim to measure the percentage of drugs bound to proteins. Equilibrium dialysis is a commonly used method where the free drug concentration at equilibrium is measured by separating the bound...

Ligand Binding Sites

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...

Ligand Binding and Linkage

Ligand Binding and Linkage

Quantitative Aspects of Drug-Receptor Interaction

Quantitative Aspects of Drug-Receptor Interaction

The receptor occupancy theory connects a drug's response to the number of occupied receptors. With higher drug concentrations, more receptors are occupied, leading to increased responses. The formation of drug-receptor complexes involves association and dissociation rates, which reach equilibrium when the forward and backward reactions are equal. The equilibrium association constant (Ka) and its inverse, the equilibrium dissociation constant (Kd), indicate drug affinity. Higher Ka and lower...

Drug-Receptor Bonds

Drug-Receptor Bonds

Drug-receptor bonds are formed through various chemical forces when drugs interact with target cells. Covalent bonds, strong and irreversible, are exemplified by DNA-alkylating anticancer agents that inhibit cell division. However, such irreversible drug binding lacks selectivity and can modify the DNA of the surrounding healthy cells. Covalent binding often contributes to tissue toxicity, as seen with chloroform and paracetamol metabolites binding to the liver, causing hepatotoxicity.
In...

You might also read

Related Articles

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

Sort by

Same author

Mechanism of AF9 (MLLT3)-Partner Dissociation in Mixed-Lineage Leukemia-Rearranged Leukemia.

JACS Au·2026

Same author

GAMA: A Robust and Automated Fragment-Based Quantum Chemistry Method for Biomolecular Systems.

The journal of physical chemistry letters·2026

Same author

Label-Free Structural Analysis of Lysophosphatidic Acid Binding to Albumin.

The journal of physical chemistry. B·2026

Same author

The Concise Guide to PHARMACOLOGY 2025/26: G protein-coupled receptors.

British journal of pharmacology·2025

Same author

MnM-W-MMGBSA: A Computational Strategy to Improve Relative Binding Free Energies of Protein-Protein Interaction Systems.

The journal of physical chemistry. B·2025

Same author

Computational and Experimental Study of the Conformational Variation of the Catalytic Residue His41 of the SARS-CoV-2 Main Protease.

The journal of physical chemistry. B·2025

Same journal

Donor-Acceptor Separation Augments Temperature Dependence of Kinetic Isotope Effects in NADH Model Hydride Transfer Reactions: Mimicking Enzyme versus Mutant Dynamics.

The journal of physical chemistry. B·2026

Same journal

Disordered Worm-Like Clusters in a Hexagonal Mesophase Former: Simulation and Thermodynamic Description.

The journal of physical chemistry. B·2026

Same journal

Comparative Biophysical Analysis of Healthy and Inflamed Intestinal Membrane Models Using Langmuir Monolayers.

The journal of physical chemistry. B·2026

Same journal

Phosphoserine Charge State Drives Ion Condensation and Spatial Polyamine Presentation in Multirepeat Silaffin.

The journal of physical chemistry. B·2026

Same journal

pH-Dependent Conformational Transition of the Glutamate-GABA Antiporter GadC Revealed by <sup>19</sup>F NMR.

The journal of physical chemistry. B·2026

Same journal

Hydrogen-Bond Network in Equimolar <i>N</i>-Methylacetamide-Water: Integrated Neutron Scattering, Molecular Dynamics, DFT-NBO-AIM, and Machine Learning Analysis.

The journal of physical chemistry. B·2026

See all related articles

Search research articles

Related Experiment Video

Updated: Jul 7, 2025

An ELISA Based Binding and Competition Method to Rapidly Determine Ligand-receptor Interactions

An ELISA Based Binding and Competition Method to Rapidly Determine Ligand-receptor Interactions

Published on: March 14, 2016

Direct Binding Methods to Measure Receptor-Ligand Interactions.

Manisha Ray¹, Aryana Sayeed¹, Madeline Ganshert¹

¹Department of Chemistry and Biochemistry, Loyola University Chicago, 1068 W. Sheridan Road, Chicago, Illinois 60660, United States.

The Journal of Physical Chemistry. B

|December 22, 2023

Summary

This summary is machine-generated.

This study introduces a novel label-free, free-solution method for measuring G-protein-coupled receptor (GPCR) interactions. This technique quantifies direct binding affinities in near-native conditions, advancing GPCR drug discovery.

More Related Videos

Real Time Measurements of Membrane Protein:Receptor Interactions Using Surface Plasmon Resonance SPR

Real Time Measurements of Membrane Protein:Receptor Interactions Using Surface Plasmon Resonance SPR

Published on: November 29, 2014

Fluorescence Biomembrane Force Probe: Concurrent Quantitation of Receptor-ligand Kinetics and Binding-induced Intracellular Signaling on a Single Cell

Fluorescence Biomembrane Force Probe: Concurrent Quantitation of Receptor-ligand Kinetics and Binding-induced Intracellular Signaling on a Single Cell

Published on: August 4, 2015

Related Experiment Videos

Last Updated: Jul 7, 2025

An ELISA Based Binding and Competition Method to Rapidly Determine Ligand-receptor Interactions

An ELISA Based Binding and Competition Method to Rapidly Determine Ligand-receptor Interactions

Published on: March 14, 2016

Real Time Measurements of Membrane Protein:Receptor Interactions Using Surface Plasmon Resonance SPR

Real Time Measurements of Membrane Protein:Receptor Interactions Using Surface Plasmon Resonance SPR

Published on: November 29, 2014

Fluorescence Biomembrane Force Probe: Concurrent Quantitation of Receptor-ligand Kinetics and Binding-induced Intracellular Signaling on a Single Cell

Fluorescence Biomembrane Force Probe: Concurrent Quantitation of Receptor-ligand Kinetics and Binding-induced Intracellular Signaling on a Single Cell

Published on: August 4, 2015

Area of Science:

Biochemistry
Pharmacology
Biophysics

Background:

G-protein-coupled receptors (GPCRs) are crucial for physiological processes, making them key drug targets.
Current indirect assays for GPCRs lack direct binding measurements, hindering drug discovery.
Existing direct methods often use labels or create non-native environments for receptors.

Purpose of the Study:

To develop a label-free, immobilization-free technique for measuring GPCR-ligand interactions.
To quantify direct binding affinities and kinetics in near-native conditions.
To overcome limitations of indirect signaling assays and traditional direct binding methods.

Main Methods:

Utilized a refractive index (RI) property-based, free-solution technique.
Measured binding interactions without receptor immobilization or labels.
Applied the method to quantify interactions between bioactive lipids and cognate lipid GPCRs in a complex cellular milieu.

Main Results:

Successfully quantified the interaction of bioactive lipids with lipid GPCRs using the developed free-solution method.
Demonstrated the ability to measure binding in near-native conditions, including other cytoplasmic lipids and proteins.
Validated a label-free approach that bypasses receptor immobilization.

Conclusions:

The developed free-solution technique offers a powerful label-free approach for studying GPCR-ligand interactions.
This method provides more physiologically relevant binding data compared to traditional assays.
Further applications across diverse receptor-ligand pairs are encouraged to explore the full potential of this technique in biomolecular interaction research.