Monitoring of receptor dimerization using plasmonic coupling of gold nanoparticles

Matthew J. Crow, Kevin Seekell, Julie H Ostrander, Adam Wax

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

The dimerization of receptors on the cell membrane is an important step in the activation of cell signaling pathways. Several methods exist for observing receptor dimerization, including coimmunoprecipitation, chemical cross-linking, and fluorescence resonance energy transfer (FRET). These techniques are limited in that only FRET is appropriate for live cells, but even that method suffers from photobleaching and bleed-through effects. In this study, we implement an alternative method for the targeting of HER-2 homodimer formation based on the plasmonic coupling of gold nanoparticles functionalized with HER-2 Ab. In the presented studies, SK-BR-3 cells, known to overexpress HER-2, are labeled with these nanoparticles and receptor colocalization is observed using plasmonic coupling. HER-2 targeted nanoparticles bound to these cells exhibit a peak resonance that is significantly red-shifted relative to those bound to similar receptors on A549 cells, which have significantly lower levels of HER-2 expression. This significant red shift indicates plasmonic coupling is occurring and points to a new avenue for assessing dimerization by monitoring their colocalization. To determine that dimerization is occurring, the refractive index of the nanoenvironment of the labels is assessed using a theoretical analysis based on the Mie coated sphere model. The results indicate scattering by single, isolated nanoparticles for the low HER-2 expressing A549 cell line, but the scattering observed for the HER-2 overexpressing SK-BR-3 cell line may only be explained by plasmonic-coupling of proximal nanoparticle pairs. To validate the conformation of nanoparticles bound to HER-2 receptors undergoing dimerization, discrete dipole approximation (DDA) models are used to assess spectra of scattering by coupled nanoparticles. Comparison of the experimental results with theoretical models indicates that NP dimers are formed for the labeling of SK-BR-3 cells, suggesting that receptor dimerization has been observed.

Original languageEnglish (US)
Pages (from-to)8532-8540
Number of pages9
JournalACS Nano
Volume5
Issue number11
DOIs
StatePublished - Nov 22 2011

Fingerprint

Dimerization
dimerization
Gold
gold
Nanoparticles
nanoparticles
Monitoring
cells
Scattering
resonance fluorescence
cultured cells
energy transfer
scattering
Cells
Cell signaling
Photobleaching
Cell membranes
red shift
Dimers
Labeling

Keywords

  • light scattering
  • nanoparticles
  • plasmonic coupling
  • receptor dimerization

Cite this

Monitoring of receptor dimerization using plasmonic coupling of gold nanoparticles. / Crow, Matthew J.; Seekell, Kevin; Ostrander, Julie H; Wax, Adam.

In: ACS Nano, Vol. 5, No. 11, 22.11.2011, p. 8532-8540.

Research output: Contribution to journalArticle

Crow, Matthew J. ; Seekell, Kevin ; Ostrander, Julie H ; Wax, Adam. / Monitoring of receptor dimerization using plasmonic coupling of gold nanoparticles. In: ACS Nano. 2011 ; Vol. 5, No. 11. pp. 8532-8540.
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AB - The dimerization of receptors on the cell membrane is an important step in the activation of cell signaling pathways. Several methods exist for observing receptor dimerization, including coimmunoprecipitation, chemical cross-linking, and fluorescence resonance energy transfer (FRET). These techniques are limited in that only FRET is appropriate for live cells, but even that method suffers from photobleaching and bleed-through effects. In this study, we implement an alternative method for the targeting of HER-2 homodimer formation based on the plasmonic coupling of gold nanoparticles functionalized with HER-2 Ab. In the presented studies, SK-BR-3 cells, known to overexpress HER-2, are labeled with these nanoparticles and receptor colocalization is observed using plasmonic coupling. HER-2 targeted nanoparticles bound to these cells exhibit a peak resonance that is significantly red-shifted relative to those bound to similar receptors on A549 cells, which have significantly lower levels of HER-2 expression. This significant red shift indicates plasmonic coupling is occurring and points to a new avenue for assessing dimerization by monitoring their colocalization. To determine that dimerization is occurring, the refractive index of the nanoenvironment of the labels is assessed using a theoretical analysis based on the Mie coated sphere model. The results indicate scattering by single, isolated nanoparticles for the low HER-2 expressing A549 cell line, but the scattering observed for the HER-2 overexpressing SK-BR-3 cell line may only be explained by plasmonic-coupling of proximal nanoparticle pairs. To validate the conformation of nanoparticles bound to HER-2 receptors undergoing dimerization, discrete dipole approximation (DDA) models are used to assess spectra of scattering by coupled nanoparticles. Comparison of the experimental results with theoretical models indicates that NP dimers are formed for the labeling of SK-BR-3 cells, suggesting that receptor dimerization has been observed.

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