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Related Experiment Videos

A "release" protocol for isothermal titration calorimetry.

H H Heerklotz1, H Binder, R M Epand

  • 1McMaster University, Health Sciences Centre, Department of Biochemistry, Hamilton, Ontario L8N 3Z5, Canada. heerklotz@ubaclu.unibas.ch

Biophysical Journal
|May 8, 1999
PubMed
Summary
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Isothermal titration calorimetry (ITC) can now model solute partitioning and ligand binding. New protocols assess solute release or ligand dissociation, providing deeper insights into molecular interactions.

Area of Science:

  • Biophysical Chemistry
  • Thermodynamics
  • Membrane Biophysics

Background:

  • Isothermal titration calorimetry (ITC) is a standard technique for studying molecular interactions.
  • ITC measures heat changes during binding or partitioning processes.
  • Existing methods have limitations in determining ligand accessibility or complex stability.

Purpose of the Study:

  • To derive a general formula for modeling ITC titration heats in binding and partitioning systems.
  • To introduce new protocols for assessing solute release or ligand dissociation.
  • To enable determination of ligand accessibility and improve accuracy of binding/partitioning parameters.

Main Methods:

  • Derivation of a general formula for ITC titration heat modeling.
  • Application of classic incorporation/binding and new release protocols.

Related Experiment Videos

  • Experimental validation using nonionic detergent C12EO7 partitioning to lipid vesicles.

    • Main Results:

    • A unified modeling approach for ITC data in binding and partitioning.
    • Demonstration of simultaneous binding/incorporation and release protocols.
    • Ability to determine if ligands access vesicle interiors within experimental timescales.

    Conclusions:

    • The new ITC modeling approach enhances the study of molecular interactions.
    • Simultaneous use of incorporation and release protocols provides critical information on ligand accessibility.
    • This method improves the accuracy of determining partition coefficients, binding constants, and transfer enthalpies.