A novel assay to probe heparin-peptide interactions using pentapeptide-stabilized gold nanoparticles.

Academic Article


  • In this article, we present a novel assay to probe the interactions between heparin and heparin-binding peptides based on CALNN pentapeptide-stabilized gold nanoparticles. This assay relies on rapid aggregation of gold nanoparticles and dramatic retardation in the presence of a large excess of heparin due to the binding of peptides to heparin. Using this method, the dissociation constant ( K d) and melting temperature ( T m) of three different peptides against heparin were determined. The results from capillary electrophoresis demonstrated that K d values measured by this method were comparatively accurate. It was found that the peptide with the lowest K d did not have the highest T m. Structural analysis by circular dichroism was performed to explain this phenomenon. A comparison with the results from affinity chromatography indicates that electrostatic interactions only are not the major determinant of the affinity between heparin and peptide, but other interactions such as hydrogen-bonding and hydrophobic interactions may play important roles in the overall interactions. This novel assay is inexpensive, label-free, and easy to implement in the laboratories, does not suffer precipitation of the heparin-peptide complex or their conformational changes caused by surface immobilization, and is expected to be a useful complement to other existing methods.
  • Authors

  • Jeong, Kyung Jae
  • Butterfield, Karen
  • Panitch, Alyssa
  • Status

    Publication Date

  • August 19, 2008
  • Published In

  • Langmuir  Journal
  • Keywords

  • Amino Acid Sequence
  • Calibration
  • Circular Dichroism
  • Gold
  • Heparin
  • Leuconostoc
  • Metal Nanoparticles
  • Molecular Sequence Data
  • Peptides
  • Transition Temperature
  • Digital Object Identifier (doi)

    Pubmed Id

  • 18646727
  • Start Page

  • 8794
  • End Page

  • 8800
  • Volume

  • 24
  • Issue

  • 16