Book contents
- Frontmatter
- Contents
- Contributors
- Preface
- LABEL-FREE BIOSENSORS
- 1 Label-free optical biosensors: An introduction
- 2 Experimental design
- 3 Extracting affinity constants from biosensor binding responses
- 4 Extracting kinetic rate constants from binding responses
- 5 Sensor surfaces and receptor deposition
- 6 Macromolecular interactions
- 7 Interactions with membranes and membrane receptors
- 8 Application of SPR technology to pharmaceutical relevant drug-receptor interactions
- 9 High-throughput analysis of biomolecular interactions and cellular responses with resonant waveguide grating biosensors
- 10 ITC-derived binding constants: Using microgram quantities of protein
- 11 Electrical impedance technology applied to cell-based assays
- Index
- Plate section
7 - Interactions with membranes and membrane receptors
Published online by Cambridge University Press: 05 May 2010
- Frontmatter
- Contents
- Contributors
- Preface
- LABEL-FREE BIOSENSORS
- 1 Label-free optical biosensors: An introduction
- 2 Experimental design
- 3 Extracting affinity constants from biosensor binding responses
- 4 Extracting kinetic rate constants from binding responses
- 5 Sensor surfaces and receptor deposition
- 6 Macromolecular interactions
- 7 Interactions with membranes and membrane receptors
- 8 Application of SPR technology to pharmaceutical relevant drug-receptor interactions
- 9 High-throughput analysis of biomolecular interactions and cellular responses with resonant waveguide grating biosensors
- 10 ITC-derived binding constants: Using microgram quantities of protein
- 11 Electrical impedance technology applied to cell-based assays
- Index
- Plate section
Summary
INTRODUCTION
Many interactions studied in the biological and biomedical sciences occur with receptors at membrane surfaces. Prominent examples are neuroreceptors, cytokine receptors, ligand-gated ion channels, G protein-coupled receptors (GPCRs), and antibody and cytokine receptors. Interactions with these receptors are especially important to academics and the pharmaceutical industry as almost half of the 100 best-selling drugs on the market are targeted to a membrane receptor. To better understand the binding mechanisms of ligands with these receptors, the ligand-receptor interactions must be probed directly in vivo or in reconstituted membrane systems. Most techniques for detailed kinetic analysis of molecular recognition events are applied in solution phase using a truncated, soluble form of the receptor. Membrane receptors, however, possess significant hydrophobic domains and are likely to have different tertiary structures and binding affinities in solution relative to those occurring in a membrane environment. This approach is limited to receptors containing a single transmembrane domain and does not allow the study of signaling cascades triggered by ligand binding to a receptor or the investigation of complex membrane proteins that often homo- or heterodimerize. However, in the last 10 years there has been significant progress in the development of techniques that allow the analysis of membrane-associated ligand–receptor interactions in a model resembling their native membrane environment.
Biophysical techniques such as patch clamping, magic angle spinning nuclear magnetic resonance (MAS-NMR), fluorescence correlation spectroscopy, fluorescence resonance energy transfer, and analytical ultracentrifugation have been applied to the analysis of binding to whole cells, membrane protoplasts, and proteoliposomes.
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- Chapter
- Information
- Label-Free BiosensorsTechniques and Applications, pp. 159 - 178Publisher: Cambridge University PressPrint publication year: 2009