title: The use of amphipols as universal molecular adapters to immobilize membrane proteins onto solid supports.
abstract: Because of the importance of their physiological functions, cell membranes represent critical targets in biological research. Membrane proteins, which make up approximately 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).
authors.affiliation: Unité Mixte de Recherche 7099, Centre National de la Recherche Scientifique and Université Paris-7, Institut de Biologie Physico-Chimique, Centre National de la Recherche Scientifique, Federated Research Institute 550, Paris, France.
authors.forename: Delphine
authors.initials: D
authors.lastname: Charvolin
authors.forename: Jean-Baptiste
authors.initials: JB
authors.lastname: Perez
authors.forename: Florent
authors.initials: F
authors.lastname: Rouvière
authors.forename: Fabrice
authors.initials: F
authors.lastname: Giusti
authors.forename: Paola
authors.initials: P
authors.lastname: Bazzacco
authors.forename: Alaa
authors.initials: A
authors.lastname: Abdine
authors.forename: Fabrice
authors.initials: F
authors.lastname: Rappaport
authors.forename: Karen L
authors.initials: KL
authors.lastname: Martinez
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Popot
mesh_terms: D000327:Adsorption; D000906:Antibodies / Q000737:chemistry; D003432:Cross-Linking Reagents / Q000737:chemistry; D008565:Membrane Proteins / Q000737:chemistry*; D008856:Microscopy, Fluorescence; D008863:Microspheres; D008954:Models, Biological; D011108:Polymers / Q000737:chemistry*; D012910:Snake Venoms / Q000737:chemistry; D020349:Surface Plasmon Resonance; D013501:Surface-Active Agents / Q000737:chemistry*
chemical_list: D000906:Antibodies; D003432:Cross-Linking Reagents; D008565:Membrane Proteins; D011108:Polymers; D012910:Snake Venoms; D013501:Surface-Active Agents