TY - JOUR
T1 - Plasmon-waveguide resonance studies of ligand binding to integral proteins in membrane fragments derived from bacterial and mammalian cells
AU - Salamon, Zdzislaw
AU - Fitch, John
AU - Cai, Minying
AU - Tumati, Suneeta
AU - Navratilova, Edita
AU - Tollin, Gordon
PY - 2009/4/1
Y1 - 2009/4/1
N2 - 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.
AB - 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.
KW - Bacterial chromatophores
KW - Cannabinoid CB1 receptor
KW - G-protein-coupled receptors
KW - Melanocortin-4 receptor
KW - Rat trigeminal ganglion
KW - Transfected HEK cells
KW - μ-Opioid receptor
UR - http://www.scopus.com/inward/record.url?scp=60549105524&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=60549105524&partnerID=8YFLogxK
U2 - 10.1016/j.ab.2009.01.019
DO - 10.1016/j.ab.2009.01.019
M3 - Article
C2 - 19454250
AN - SCOPUS:60549105524
SN - 0003-2697
VL - 387
SP - 95
EP - 101
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 1
ER -