Plasmon-waveguide resonance studies of ligand binding to integral proteins in membrane fragments derived from bacterial and mammalian cells

Zdzislaw Salamon, John Fitch, Minying Cai, Suneeta Tumati, Edita Navratilova, Gordon Tollin

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

A procedure has been developed for directly depositing membrane fragments derived from bacterial cells (chromatophores from Rhodopseudomonas sphaeroides) and mammalian cells (μ-opioid receptor- and MC4 receptor-transfected human embryonic kidney (HEK) cells and rat trigeminal ganglion cells) on the silica surface of a plasmon-waveguide resonance (PWR) spectrometer. Binding of ligands (cytochrome c2 for the chromatophores, the peptide agonists DAMGO and melanotan-II that are specific for the μ-opioid and MC4 receptors, and two nonpeptide agonists that are specific for the CB1 receptor) to these membrane fragments has been observed and characterized with high sensitivity using PWR spectral shifts. The KD values obtained are in excellent agreement with conventional pharmacological assays and with prior PWR studies using purified receptors inserted into deposited lipid bilayer membranes. These studies provide a new tool for obtaining useful biological information about receptor-mediated processes in real biological membranes.

Original languageEnglish (US)
Pages (from-to)95-101
Number of pages7
JournalAnalytical Biochemistry
Volume387
Issue number1
DOIs
StatePublished - Apr 1 2009

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Keywords

  • Bacterial chromatophores
  • Cannabinoid CB1 receptor
  • G-protein-coupled receptors
  • Melanocortin-4 receptor
  • Rat trigeminal ganglion
  • Transfected HEK cells
  • μ-Opioid receptor

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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