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

Factors Affecting Protein-Drug Binding: Protein-Related Factors01:20

Factors Affecting Protein-Drug Binding: Protein-Related Factors

576
Drug binding to proteins is a key aspect of pharmacokinetics and can influence a drug's distribution, absorption, and elimination in the body. Several factors, including the drug's physiochemical properties, protein concentration, disease states, and the number of binding sites on the protein, influence this process.
The physicochemical properties of a drug play a significant role in its ability to bind to proteins. Lipophilic drugs, which dissolve in fats, oils, and lipids, can be...
576
The Equilibrium Binding Constant and Binding Strength02:18

The Equilibrium Binding Constant and Binding Strength

15.2K
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:
15.2K
Protein-protein Interfaces02:04

Protein-protein Interfaces

14.8K
Many proteins form complexes to carry out their functions, making protein-protein interactions (PPIs) essential for an organism's survival. Most PPIs are stabilized by numerous weak noncovalent chemical forces. The physical shape of the interfaces determines the way two proteins interact. Many globular proteins have closely-matching shapes on their surfaces, which form a large number of weak bonds. Additionally, many PPIs occur between two helices or between a surface cleft and a...
14.8K
Yeast Signaling01:28

Yeast Signaling

17.3K
Yeasts are single-celled organisms, but unlike bacteria, they are eukaryotes (cells with a nucleus). Cell signaling in yeast is similar to signaling in other eukaryotic cells. A ligand, such as a protein or a small molecule released from a yeast cell, attaches to a receptor on the cell surface. The binding stimulates second-messenger kinases to activate or inactivate transcription factors that further regulate gene expression. Many of the yeast intracellular signaling cascades have similar...
17.3K
Drug Distribution: Plasma Protein Binding01:29

Drug Distribution: Plasma Protein Binding

9.0K
Drugs predominantly attach to plasma proteins, with only a small percentage remaining unbound. The unbound portion can be calculated as one minus the bound fraction. Acidic drugs form large, inactive complexes by reversibly binding to plasma albumin, which prevents them from diffusing across biological barriers. These drug-protein complexes act as reservoirs for the drugs. As the concentration of unbound drugs decreases, these complexes quickly dissociate to release the free drug, maintaining...
9.0K
Protein-Drug Binding: Determination Methods01:22

Protein-Drug Binding: Determination Methods

676
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...
676

You might also read

Related Articles

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

Sort by
Same author

Stringent selection drives convergence toward omicron-like SARS-CoV-2 receptor-binding motifs.

Nature communications·2026
Same author

Electrostatic interactions constrain generalization of porous-media models for intracellular diffusion in mammalian cells.

Proceedings of the National Academy of Sciences of the United States of America·2026
Same author

Reversing protonation of weakly basic drugs greatly enhances intracellular diffusion and decreases lysosomal sequestration.

eLife·2024
Same author

Promiscuous Janus kinase binding to cytokine receptors modulates signaling efficiencies and contributes to cytokine pleiotropy.

Science signaling·2024
Same author

Variant-proof high affinity ACE2 antagonist limits SARS-CoV-2 replication in upper and lower airways.

Nature communications·2024
Same author

Harnessing the power of IFN for therapeutic approaches to COVID-19.

Journal of virology·2024

Related Experiment Video

Updated: Feb 11, 2026

Primer-Free Aptamer Selection Using A Random DNA Library
11:14

Primer-Free Aptamer Selection Using A Random DNA Library

Published on: July 26, 2010

25.4K

Selecting for Fast Protein-Protein Association As Demonstrated on a Random TEM1 Yeast Library Binding BLIP.

Ruth Cohen-Khait1, Gideon Schreiber1

  • 1Department of Biomolecular Sciences , Weizmann Institute of Science , 76100 Rehovot , Israel.

Biochemistry
|April 20, 2018
PubMed
Summary

This study introduces a novel method for selecting protein variants with faster binding rates, crucial for cellular processes and drug development. The technique refines selection conditions to specifically isolate proteins with enhanced association kinetics.

More Related Videos

Creating Highly Specific Chemically Induced Protein Dimerization Systems by Stepwise Phage Selection of a Combinatorial Single-Domain Antibody Library
10:17

Creating Highly Specific Chemically Induced Protein Dimerization Systems by Stepwise Phage Selection of a Combinatorial Single-Domain Antibody Library

Published on: January 14, 2020

8.3K
A Yeast 2-Hybrid Screen in Batch to Compare Protein Interactions
14:23

A Yeast 2-Hybrid Screen in Batch to Compare Protein Interactions

Published on: June 6, 2018

14.2K

Related Experiment Videos

Last Updated: Feb 11, 2026

Primer-Free Aptamer Selection Using A Random DNA Library
11:14

Primer-Free Aptamer Selection Using A Random DNA Library

Published on: July 26, 2010

25.4K
Creating Highly Specific Chemically Induced Protein Dimerization Systems by Stepwise Phage Selection of a Combinatorial Single-Domain Antibody Library
10:17

Creating Highly Specific Chemically Induced Protein Dimerization Systems by Stepwise Phage Selection of a Combinatorial Single-Domain Antibody Library

Published on: January 14, 2020

8.3K
A Yeast 2-Hybrid Screen in Batch to Compare Protein Interactions
14:23

A Yeast 2-Hybrid Screen in Batch to Compare Protein Interactions

Published on: June 6, 2018

14.2K

Area of Science:

  • Biochemistry and Molecular Biology
  • Cellular Biology
  • Biotechnology

Background:

  • Protein-protein interactions are fundamental to cellular functions, but in vivo conditions may prevent equilibrium.
  • In vitro thermodynamic affinity alone may not fully represent in vivo protein interactions.
  • Binding kinetics, including association and dissociation rates, are critical for protein activity within the cell.

Purpose of the Study:

  • To develop a method for selecting protein variants with enhanced association rate constants (kon).
  • To refine selection conditions for isolating fast-associating protein variants from a library.
  • To explore the utility of pre-equilibrium selection for identifying proteins with improved binding kinetics.

Main Methods:

  • Utilized a pre-equilibrium selection strategy on a large random library of TEM1-β-lactamase variants.
  • Optimized selection conditions, including ligand concentration and incubation time, against β-lactamase inhibitor protein (BLIP).
  • Employed a secondary sorting step to improve the enrichment of desired variants and reduce noise.

Main Results:

  • Successfully developed a method to specifically select for protein variants with faster association rate constants.
  • Demonstrated that optimal selection depends on ligand concentration and incubation duration.
  • Showed that a second round of selection significantly increases the proportion of fast-associating variants.

Conclusions:

  • The developed pre-equilibrium selection method effectively identifies protein variants with enhanced association kinetics.
  • Fast-associating protein variants have significant implications for drug discovery and biotechnological applications.
  • This approach offers a valuable tool for engineering proteins with improved binding dynamics for therapeutic and industrial purposes.