Binding assays with artificial tethered membranes using surface plasmon resonance.
Surface sensitive optical techniques based on surface plasmon resonance have become interesting for biosciences in the context of biorecognition and binding studies at functional surfaces. We use surface plasmon resonance spectroscopy (SPS) in combination with surface plasmon enhanced fluorescence spectroscopy (SPFS) for the characterization of interaction processes associated with biomembranes. The biological membrane is mimicked by a tethered membrane consisting of a planar lipid bilayer attached to a gold surface via a hydrophilic anchor peptide. The interaction between membrane-bound hydrophobic compounds and free hydrophilic molecules is monitored in real-time and with high sensitivity and selectivity by combined SPS/SPFS. In this review we shortly discuss the principles of surface plasmon resonance and its utilization in SPS and SPFS. A detailed description of the required instrumentation for combined SPS and SPFS is presented. Furthermore, we outline the design of a binding assay with a tethered bilayer and the procedure of the artificial membrane system built-up is delineated. We also present examples that demonstrate the potential of combined SPS/SPFS assays with artificial tethered membranes. The method provides insight into the interaction of integral membrane proteins with various hydrophilic ligands and the specific recognition of small lipophilic molecules by soluble proteins.
Wiltschi
Birgit
B
Max Planck Institute for Biochemistry, Department of Membrane Biochemistry, D-82152 Martinsried, Germany. wiltschi@biochem.mpg.de
Knoll
Wolfgang
W
Sinner
Eva-Kathrin
EK
D000595:Amino Acid Sequence; D002232:Carbocyanines / Q000737:chemistry; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry; D007700:Kinetics; D008051:Lipid Bilayers; D008565:Membrane Proteins / Q000378:metabolism; D008567:Membranes, Artificial; D008969:Molecular Sequence Data; D012828:Silicones; D020349:Surface Plasmon Resonance / Q000295:instrumentation / Q000379:methods*
D002232:Carbocyanines; D005456:Fluorescent Dyes; D008051:Lipid Bilayers; D008565:Membrane Proteins; D008567:Membranes, Artificial; D012828:Silicones; C085321:cyanine dye 5