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

Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

1.9K
Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
1.9K

You might also read

Related Articles

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

Sort by
Same author

Aromatic ring transforms DNA binding modes of an octahedral rhodium complex.

Biophysical journal·2026
Same author

Bacteriophage T4 gene 32 protein: Insights into its interaction with ssDNA, binding cooperativity, and conformational change.

PloS one·2026
Same author

Mechanism of SARS-CoV-2 Nucleocapsid Protein Phosphorylation-Induced Functional Switch.

Viruses·2026
Same author

Bacteriophage T4 gene 32 protein: Insights into its Interaction with ssDNA, binding cooperativity, and conformational change.

bioRxiv : the preprint server for biology·2025
Same author

Engineering metal-oxide interfaces via controlled exsolution in Ir-doped Y<sub>2</sub>Ru<sub>2</sub>O<sub>7</sub> pyrochlore for superior hydrogen evolution reaction.

Journal of colloid and interface science·2025
Same author

Single-molecule measurements of double-stranded DNA condensation.

Biophysical journal·2025

Related Experiment Video

Updated: May 1, 2026

Direct Observation of Enzymes Replicating DNA Using a Single-molecule DNA Stretching Assay
17:03

Direct Observation of Enzymes Replicating DNA Using a Single-molecule DNA Stretching Assay

Published on: March 24, 2010

18.2K

Quantifying DNA-protein interactions by single molecule stretching.

Mark C Williams1, Ioulia Rouzina, Richard L Karpel

  • 1Department of Physics, 111 Dana Research Center, Northeastern University, Boston, Massachusetts 02115, USA.

Methods in Cell Biology
|October 30, 2007
PubMed
Summary
This summary is machine-generated.

This study introduces a novel method to measure DNA-protein interactions by stretching DNA to quantify binding free energy. The technique accurately determined binding constants for bacteriophage T4 gene 32 protein (gp32).

More Related Videos

Combining Single-molecule Manipulation and Imaging for the Study of Protein-DNA Interactions
14:43

Combining Single-molecule Manipulation and Imaging for the Study of Protein-DNA Interactions

Published on: August 27, 2014

10.9K
Analyzing DNA-Protein Interactions with Streptavidin-Based Biolayer Interferometry
08:07

Analyzing DNA-Protein Interactions with Streptavidin-Based Biolayer Interferometry

Published on: January 17, 2025

1.9K

Related Experiment Videos

Last Updated: May 1, 2026

Direct Observation of Enzymes Replicating DNA Using a Single-molecule DNA Stretching Assay
17:03

Direct Observation of Enzymes Replicating DNA Using a Single-molecule DNA Stretching Assay

Published on: March 24, 2010

18.2K
Combining Single-molecule Manipulation and Imaging for the Study of Protein-DNA Interactions
14:43

Combining Single-molecule Manipulation and Imaging for the Study of Protein-DNA Interactions

Published on: August 27, 2014

10.9K
Analyzing DNA-Protein Interactions with Streptavidin-Based Biolayer Interferometry
08:07

Analyzing DNA-Protein Interactions with Streptavidin-Based Biolayer Interferometry

Published on: January 17, 2025

1.9K

Area of Science:

  • Biophysics
  • Molecular Biology
  • Biochemistry

Background:

  • Quantifying DNA-protein interactions is crucial for understanding biological processes.
  • Existing methods may have limitations in precision or scope.
  • Force-induced DNA melting offers a new approach to probe these interactions.

Purpose of the Study:

  • To develop and validate a new method for quantifying DNA-protein interactions using force-induced DNA melting.
  • To determine the binding free energy and equilibrium binding constants of DNA-binding proteins.
  • To investigate the binding characteristics of bacteriophage T4 gene 32 protein (gp32) to single-stranded DNA (ssDNA).

Main Methods:

  • Stretching a single double-stranded DNA (dsDNA) molecule beyond its contour length to induce melting into single-stranded DNA (ssDNA).
  • Applying this force-induced DNA melting in solutions containing DNA-binding ligands, specifically proteins.
  • Measuring the dependence of melting force on protein concentration to derive equilibrium binding constants.

Main Results:

  • The force-induced DNA melting method successfully quantified the free energy of protein binding.
  • Equilibrium binding constants for bacteriophage T4 gene 32 protein (gp32) to ssDNA were obtained under various solution conditions.
  • Analysis revealed a salt-dependent conformational change regulating gp32 DNA binding near physiological salt concentrations.

Conclusions:

  • Force-induced DNA melting is a viable and effective method for quantifying DNA-protein interactions.
  • The study provides new insights into the binding mechanisms of gp32, highlighting the role of salt concentration.
  • This technique has broad applicability for studying various DNA-binding ligands and proteins.