期刊
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
卷 106, 期 2, 页码 405-410出版社
NATL ACAD SCIENCES
DOI: 10.1073/pnas.0807132106
关键词
diagnostics; drug discovery; immobilization; chips bioreactors
资金
- Centre National de la Recherche Scientifique
- University Paris-7
- European Union [LSHG-CT-2005-513770]
- Centre National de la Recherche Scientifique [ST 83747-04]
- French Ministry of Research [ANR06-EMPB-027]
- Danish Research Council [FNU 272-05-0355, NABIIT 2106-05-0036]
- French Embassy in Denmark
- Marie Curie Early Stage Training Fellowship
- European Commission Network BioMem
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).
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