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Related Concept Videos

Allosteric Regulation01:08

Allosteric Regulation

Allosteric regulation of enzymes occurs when the binding of an effector molecule to a site that is different from the active site causes a change in the enzymatic activity. This alternate site is called an allosteric site, and an enzyme can contain more than one of these sites. Allosteric regulation can either be positive or negative, resulting in an increase or decrease in enzyme activity. Most enzymes that display allosteric regulation are metabolic enzymes involved in the degradation or...
Allosteric Regulation01:08

Allosteric Regulation

Allosteric regulation of enzymes occurs when the binding of an effector molecule to a site that is different from the active site causes a change in the enzymatic activity. This alternate site is called an allosteric site, and an enzyme can contain more than one of these sites. Allosteric regulation can either be positive or negative, resulting in an increase or decrease in enzyme activity. Most enzymes that display allosteric regulation are metabolic enzymes involved in the degradation or...
Cooperative Allosteric Transitions01:58

Cooperative Allosteric Transitions

Cooperative allosteric transitions can occur in multimeric proteins, where each subunit of the protein has its own ligand-binding site. When a ligand binds to any of these subunits, it triggers a conformational change that affects the binding sites in the other subunits; this can change the affinity of the other sites for their respective ligands. The ability of the protein to change the shape of its binding site is attributed to the presence of a mix of flexible and stable segments in the...
Cooperative Allosteric Transitions01:58

Cooperative Allosteric Transitions

Cooperative allosteric transitions can occur in multimeric proteins, where each subunit of the protein has its own ligand-binding site. When a ligand binds to any of these subunits, it triggers a conformational change that affects the binding sites in the other subunits; this can change the affinity of the other sites for their respective ligands. The ability of the protein to change the shape of its binding site is attributed to the presence of a mix of flexible and stable segments in the...
Cooperative Allosteric Transitions01:58

Cooperative Allosteric Transitions

Cooperative allosteric transitions can occur in multimeric proteins, where each subunit of the protein has its own ligand-binding site. When a ligand binds to any of these subunits, it triggers a conformational change that affects the binding sites in the other subunits; this can change the affinity of the other sites for their respective ligands. The ability of the protein to change the shape of its binding site is attributed to the presence of a mix of flexible and stable segments in the...
Allosteric Proteins-ATCase01:19

Allosteric Proteins-ATCase

Binding sites linkages can regulate a protein's function.  For example, enzyme activity is often regulated through a feedback mechanism where the end product of the biochemical process serves as an inhibitor.
Aspartate transcarbamoylase (ATCase) is a cytosolic enzyme that catalyzes the condensation of L-aspartate and carbamoyl phosphate to  N-carbamoyl-L-aspartate. This reaction is the first step in pyrimidine biosynthesis. UTP and CTP, the end products of the pyrimidine synthesis pathway,...

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Related Experiment Video

Updated: May 27, 2026

Bio-layer Interferometry for Measuring Kinetics of Protein-protein Interactions and Allosteric Ligand Effects
13:57

Bio-layer Interferometry for Measuring Kinetics of Protein-protein Interactions and Allosteric Ligand Effects

Published on: February 18, 2014

Detecting "silent" allosteric coupling.

Harvey F Fisher1

  • 1Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, KS, USA. hfisher@kumc.edu

Methods in Molecular Biology (Clifton, N.J.)
|November 5, 2011
PubMed
Summary
This summary is machine-generated.

Isothermal titration calorimetry reveals "isoergonic cooperativity" or "silent coupling" in ligand binding, where thermodynamic parameters beyond binding free energy (ΔG°) explain complex allosteric behavior. This method offers precise insights into molecular interactions.

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Last Updated: May 27, 2026

Bio-layer Interferometry for Measuring Kinetics of Protein-protein Interactions and Allosteric Ligand Effects
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Use of Label-free Optical Biosensors to Detect Modulation of Potassium Channels by G-protein Coupled Receptors
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Use of Label-free Optical Biosensors to Detect Modulation of Potassium Channels by G-protein Coupled Receptors

Published on: February 10, 2014

Area of Science:

  • Biophysical Chemistry
  • Biochemistry
  • Molecular Biology

Background:

  • Allosteric regulation is crucial for biological processes.
  • Previous studies often focused solely on binding free energy (ΔG°), potentially missing complex thermodynamic behaviors.

Purpose of the Study:

  • To investigate allosteric ligand binding beyond ΔG° using isothermal titration calorimetry (ITC).
  • To explain phenomena termed "isoergonic cooperativity" or "silent coupling" through comprehensive thermodynamic analysis.

Main Methods:

  • Isothermal titration calorimetry (ITC) was employed to directly measure thermodynamic parameters.
  • Analysis extended beyond ΔG° to include ΔH°, ΔS°, ΔC(p)°, and d(ΔC(p)°/dt).
  • Basic linkage theory was applied to interpret binding events.

Main Results:

  • Observed cases where allosteric activators lacked ΔG° allostery but exhibited entropy-compensated enthalpy changes.
  • Demonstrated that comprehensive thermodynamic data explains "isoergonic cooperativity" and "silent coupling."
  • Highlighted the limitations of van't Hoff and Arrhenius plots for studying allosteric phenomena.

Conclusions:

  • Direct calorimetric determination of multiple thermodynamic parameters provides a more complete understanding of ligand binding than ΔG° alone.
  • ITC offers superior precision and depth of analysis compared to van't Hoff and Arrhenius methods for studying allosteric effects.
  • Isoergonic cooperativity is explained by considering enthalpy-entropy compensation and linkage theory beyond simple free energy changes.