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Related Concept Videos

SDS-PAGE01:27

SDS-PAGE

Gel electrophoresis is a method that separates biological macromolecules like nucleic acids or proteins by forcing them to pass through a gel matrix under an electric field.
A variation of gel electrophoresis, termed  polyacrylamide gel electrophoresis (PAGE), is commonly used for separating proteins according to their molecular size by passing them through a polyacrylamide gel. Because of the varying charges associated with amino acid side chains, PAGE can be used to separate intact proteins...

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Single-Molecule Real-Time Visualization of DNA Unwinding by CMG Helicase
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Single-Molecule Real-Time Visualization of DNA Unwinding by CMG Helicase

Published on: September 27, 2024

Visualization of different pathways of DNA release from interpolyelectrolyte complex.

Mehran Asad Ayoubi1, Anatoly A Zinchenko, Olga E Philippova

  • 1Physics Department, Moscow State University, Moscow 119992, Russia, and Department of Physics, Graduate School of Science, Kyoto University, Kyoto 606-8502, Japan.

The Journal of Physical Chemistry. B
|April 17, 2007
PubMed
Summary

Double-stranded DNA release from poly(allylamine hydrochloride) complexes was observed. DNA release pathways depend on salt concentration, forming intermediate structures at low salt and direct release at high salt.

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Combining Single-molecule Manipulation and Imaging for the Study of Protein-DNA Interactions
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Single-Molecule Real-Time Visualization of DNA Unwinding by CMG Helicase
07:37

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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

Area of Science:

  • Polymer Science
  • Biophysics
  • Materials Science

Background:

  • Interpolyelectrolyte complexes (IPECs) are formed between oppositely charged polymers.
  • Understanding the release mechanisms of DNA from IPECs is crucial for applications in gene delivery and biomaterials.
  • Poly(allylamine hydrochloride) (PAH) and poly(sodium styrenesulfonate) (PSS) are commonly used polyelectrolytes.

Purpose of the Study:

  • To directly observe the release of double-stranded DNA (dsDNA) from its complex with PAH.
  • To investigate the influence of added salt concentration on the DNA release pathway.
  • To elucidate the structural transitions during DNA release from PAH/dsDNA complexes.

Main Methods:

  • Formation of PAH/dsDNA interpolyelectrolyte complexes.
  • Exchange reaction using added polyanion, poly(sodium styrenesulfonate) (PSS).
  • Direct observation of DNA release using fluorescence microscopy.
  • Systematic variation of low-molecular-weight salt concentration.

Main Results:

  • DNA release was directly visualized by fluorescence microscopy.
  • The pathway of DNA release is critically dependent on the concentration of added salt.
  • At low salt concentrations, an intermediate "beads-on-string" structure was observed during DNA release.
  • At high salt concentrations, DNA was released directly from a globule to coil state without intermediate structures.

Conclusions:

  • The salt concentration dictates the mechanism of DNA release from PAH/dsDNA complexes.
  • The "beads-on-string" intermediate is a key feature at low salt, suggesting a stepwise dissociation process.
  • High salt conditions promote a more direct and complete dissociation of the DNA from the complex.