Advances in membrane receptor screening and analysis.
During the last decade there has been significant progress in the development of analytical techniques for the screening of ligand binding to membranes and membrane receptors. This review focuses on developments using label-free assays that facilitate ligand-membrane-receptor screening without the need for chemical-, biological- or radiological-labelled reagents. These assays include acoustic, optical surface plasmon resonance biosensing, sedimentation (analytical ultracentrifugation), chromatographic assays, isothermal titration calorimetry and differential scanning calorimetry. The merits and applications of cell-based screening systems and of different model membrane systems, including planar supported lipid layers, bead-supported membranes and lipid micro-arrays, are discussed. Recent advances involving more established techniques including intrinsic fluorescence, FRET spectroscopy, scintillation proximity assays and automated patch clamping are presented along with applications to peripheral membrane proteins, ion channels and G protein-coupled receptors. Novel high-throughput assays for determination of drug- and protein-partitioning in membranes are also highlighted. To aid the experimenter, a brief synopsis of the techniques commonly employed to purify and reconstitute membranes and membrane receptors is included.
Cooper
Matthew A
MA
Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, UK. mc221@cam.ac.uk
D000818:Animals; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods*; D002152:Calorimetry, Differential Scanning / Q000379:methods; D006801:Humans; D008567:Membranes, Artificial; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D020349:Surface Plasmon Resonance / Q000295:instrumentation / Q000379:methods; D014461:Ultracentrifugation / Q000379:methods
D008567:Membranes, Artificial; D011956:Receptors, Cell Surface