The use of amphipols as universal molecular adapters to immobilize membrane proteins onto solid supports
- 13 January 2009
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences of the United States of America
- Vol. 106 (2), 405-410
- https://doi.org/10.1073/pnas.0807132106
Abstract
Because of the importance of their physiological functions, cell membranes represent critical targets in biological research. Membrane proteins, which make up approximate to 1/3 of the proteome, interact with a wide range of small ligands and macromolecular partners as well as with foreign molecules such as synthetic drugs, antibodies, toxins, or surface recognition proteins of pathogenic organisms. Whether it is for the sake of basic biomedical or pharmacological research, it is of great interest to develop tools facilitating the study of these interactions. Surface-based in vitro assays are appealing because they require minimum quantities of reagents, and they are suitable for multiplexing and high-throughput screening. We introduce here a general method for immobilizing functional, unmodified integral membrane proteins onto solid supports, thanks to amphipathic polymers called "amphipols.'' The key point of this approach is that functionalized amphipols can be used as universal adapters to associate any membrane protein to virtually any kind of support while stabilizing its native state. The generality and versatility of this strategy is demonstrated by using 5 different target proteins, 2 types of supports (chips and beads), 2 types of ligands (antibodies and a snake toxin), and 2 detection methods (surface plasmon resonance and fluorescence microscopy).Keywords
This publication has 36 references indexed in Scilit:
- Bacteriorhodopsin/Amphipol Complexes: Structural and Functional PropertiesBiophysical Journal, 2008
- PDZ Domain Binding Selectivity Is Optimized Across the Mouse ProteomeScience, 2007
- Specifically Immobilised Aldo/Keto Reductase AKR1A1 Shows a Dramatic Increase in Activity Relative to the Randomly Immobilised EnzymeChemBioChem, 2007
- Functional GPCR MicroarraysJournal of the American Chemical Society, 2005
- Ligand Binding to G Protein-Coupled Receptors in Tethered Cell MembranesLangmuir, 2003
- Protein chip technologyCurrent Opinion in Chemical Biology, 2002
- Global Analysis of Protein Activities Using Proteome ChipsScience, 2001
- Association between Hydrophobically Modified Polyanions and Negatively Charged Bovine Serum AlbuminThe Journal of Physical Chemistry B, 1998
- Radiolabeled .alpha.-bungarotoxin derivatives: kinetic interaction with nicotinic acetylcholine receptorsBiochemistry, 1981
- Studies of the binding of α‐bungarotoxin to membrane‐bound and detergent‐dispersed acetylcholine receptors from Torpedo electric tissueFEBS Letters, 1972