Detection of channel proximity by nanoparticle-assisted delaying of toxin binding; a combined patch-clamp and flow cytometric energy transfer study

Bálint Rubovszky, Péter Hajdú, Z. Krasznai, R. Gáspár, Thomas A. Waldmann, S. Damjanovich, L. Bene

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Gold nanoparticles of 30 nm diameter bound to cell-surface receptor major histocompatibility complex glycoproteins (MHCI and MHCII), interleukin-2 receptor α subunit (IL-2Rα), very late antigen-4 (VLA-4) integrin, transferrin receptor, and the receptor-type protein tyrosin phosphatase CD45 are shown by the patch-clamp technique to selectively modulate binding characteristics of Pi2 toxin, an efficient blocker of K v1.3 channels. After correlating the electrophysiological data with those on the underlying receptor clusters obtained by simultaneously conducted flow cytometric energy transfer measurements, the modulation was proved to be sensitive to the density and size of the receptor clusters, and to the locations of the receptors as well. Based on the observation that engagement of MHCII by a monoclonal antibody down-regulates channel current and based on the close nanometer-scale proximity of the MHCI and MHCII glycoproteins, an analogous experiment was carried out when gold nanoparticles bound to MHCI delayed down-regulation of the Kv1.3 current initiated by ligation of MHCII. Localization of Kv1.3 channels in the nanometer-scale vicinity of the MHC-containing lipid rafts is demonstrated for the first time. A method is proposed for detecting receptor-channel or receptor-receptor proximity by observing nanoparticle-induced increase in relaxation times following concentration jumps of ligands binding to channels or to receptors capable of regulating channel currents.

Original languageEnglish
Pages (from-to)127-143
Number of pages17
JournalEuropean Biophysics Journal
Volume34
Issue number2
DOIs
Publication statusPublished - Mar 2005

Fingerprint

Energy Transfer
Nanoparticles
Gold
Glycoproteins
Down-Regulation
Integrin alpha4beta1
Transferrin Receptors
Interleukin-2 Receptors
Phosphoprotein Phosphatases
Cell Surface Receptors
Patch-Clamp Techniques
Major Histocompatibility Complex
Integrins
Ligation
Monoclonal Antibodies
Ligands
Lipids

Keywords

  • Interleukin receptor
  • Lipid raft
  • Major histocompatibility complex glycoproteins
  • Potassium channel
  • Very late antigen integrin

ASJC Scopus subject areas

  • Biophysics

Cite this

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title = "Detection of channel proximity by nanoparticle-assisted delaying of toxin binding; a combined patch-clamp and flow cytometric energy transfer study",
abstract = "Gold nanoparticles of 30 nm diameter bound to cell-surface receptor major histocompatibility complex glycoproteins (MHCI and MHCII), interleukin-2 receptor α subunit (IL-2Rα), very late antigen-4 (VLA-4) integrin, transferrin receptor, and the receptor-type protein tyrosin phosphatase CD45 are shown by the patch-clamp technique to selectively modulate binding characteristics of Pi2 toxin, an efficient blocker of K v1.3 channels. After correlating the electrophysiological data with those on the underlying receptor clusters obtained by simultaneously conducted flow cytometric energy transfer measurements, the modulation was proved to be sensitive to the density and size of the receptor clusters, and to the locations of the receptors as well. Based on the observation that engagement of MHCII by a monoclonal antibody down-regulates channel current and based on the close nanometer-scale proximity of the MHCI and MHCII glycoproteins, an analogous experiment was carried out when gold nanoparticles bound to MHCI delayed down-regulation of the Kv1.3 current initiated by ligation of MHCII. Localization of Kv1.3 channels in the nanometer-scale vicinity of the MHC-containing lipid rafts is demonstrated for the first time. A method is proposed for detecting receptor-channel or receptor-receptor proximity by observing nanoparticle-induced increase in relaxation times following concentration jumps of ligands binding to channels or to receptors capable of regulating channel currents.",
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author = "B{\'a}lint Rubovszky and P{\'e}ter Hajd{\'u} and Z. Krasznai and R. G{\'a}sp{\'a}r and Waldmann, {Thomas A.} and S. Damjanovich and L. Bene",
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T1 - Detection of channel proximity by nanoparticle-assisted delaying of toxin binding; a combined patch-clamp and flow cytometric energy transfer study

AU - Rubovszky, Bálint

AU - Hajdú, Péter

AU - Krasznai, Z.

AU - Gáspár, R.

AU - Waldmann, Thomas A.

AU - Damjanovich, S.

AU - Bene, L.

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