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Related Concept Videos

Single-Strand DNA Binding Proteins01:03

Single-Strand DNA Binding Proteins

For successful DNA replication, the unwinding of double-stranded DNA must be accompanied by stabilization and protection of the separated single strands of the DNA. This crucial task is performed by single-strand DNA-binding (SSB) proteins. They bind to the DNA in a sequence-independent manner, which means that the nitrogenous bases of the DNA need not be present in a specific order for binding of SSB proteins to it. The binding of SSB proteins straightens single-stranded DNA (ssDNA) and makes...

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Related Experiment Video

Updated: May 18, 2026

Determining the Thermodynamic and Kinetic Association of a DNA Aptamer and Tetracycline Using Isothermal Titration Calorimetry
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Determining the Thermodynamic and Kinetic Association of a DNA Aptamer and Tetracycline Using Isothermal Titration Calorimetry

Published on: August 23, 2022

SSB binding to ssDNA using isothermal titration calorimetry.

Alexander G Kozlov1, Timothy M Lohman

  • 1Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, MO, USA.

Methods in Molecular Biology (Clifton, N.J.)
|September 15, 2012
PubMed
Summary

Isothermal titration calorimetry (ITC) quantifies protein-DNA interactions by measuring binding energetics and stoichiometry. This thermodynamic data is crucial for understanding interaction stability and function, especially for oligomeric proteins like SSB.

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Determining the Thermodynamic and Kinetic Association of a DNA Aptamer and Tetracycline Using Isothermal Titration Calorimetry
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Collecting Variable-concentration Isothermal Titration Calorimetry Datasets in Order to Determine Binding Mechanisms

Published on: April 7, 2011

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Thermodynamics

Background:

  • Isothermal titration calorimetry (ITC) is a key technique for analyzing molecular interactions in solution.
  • Understanding protein-DNA interactions is vital for elucidating biological stability, specificity, and function.
  • Oligomeric proteins, such as single-stranded DNA binding proteins (SSBs), exhibit complex binding behaviors.

Purpose of the Study:

  • To detail the procedures for performing and analyzing Isothermal Titration Calorimetry (ITC) studies.
  • To highlight the quantitative thermodynamic information obtainable from ITC, including stoichiometry and binding energetics.
  • To emphasize the influence of solution conditions on the binding modes of oligomeric proteins, using E. coli and D. radiodurans SSB as examples.

Main Methods:

  • Isothermal titration calorimetry (ITC) was employed to measure binding thermodynamics.
  • Analysis of binding data to determine stoichiometry, affinity, enthalpy, and entropy changes.
  • Comparative study of two different SSB proteins (E. coli and D. radiodurans) to illustrate oligomeric protein binding.

Main Results:

  • ITC successfully provided quantitative data on stoichiometries and binding energetics for protein-DNA interactions.
  • The study demonstrated the ability of ITC to reveal site-site interactions and cooperativity.
  • Differences in oligomeric structure (tetramer vs. dimer) influenced SSB binding modes under varying solution conditions.

Conclusions:

  • ITC is a powerful and versatile method for a comprehensive thermodynamic characterization of protein-DNA interactions.
  • Accurate thermodynamic data from ITC is essential for understanding the stability and specificity of these interactions.
  • Solution conditions significantly impact the binding mechanisms of oligomeric proteins, necessitating careful experimental design and interpretation.