The use of amphipols as universal molecular adapters to immobilize membrane proteins onto solid supports.
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).
Charvolin
Delphine
D
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.
Perez
Jean-Baptiste
JB
Rouvière
Florent
F
Giusti
Fabrice
F
Bazzacco
Paola
P
Abdine
Alaa
A
Rappaport
Fabrice
F
Martinez
Karen L
KL
Popot
Jean-Luc
JL
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*
D000906:Antibodies; D003432:Cross-Linking Reagents; D008565:Membrane Proteins; D011108:Polymers; D012910:Snake Venoms; D013501:Surface-Active Agents