The use of amphipols as universal molecular adapters to immobilize membrane proteins onto solid supports
| Autor: | Florent Rouvière, Delphine Charvolin, Jean-Luc Popot, Jean-Baptiste Perez, Fabrice Rappaport, Karen L. Martinez, Fabrice Giusti, Paola Bazzacco, Alaa Abdine |
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| Přispěvatelé: | Laboratoire d'Informatique, de Modélisation et d'Optimisation des Systèmes (LIMOS), Ecole Nationale Supérieure des Mines de St Etienne-Université Clermont Auvergne [2017-2020] (UCA [2017-2020])-Centre National de la Recherche Scientifique (CNRS), Physiologie membranaire et moléculaire du chloroplaste (PMMC), Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS), Ecole Nationale Supérieure des Mines de St Etienne (ENSM ST-ETIENNE)-Université Clermont Auvergne [2017-2020] (UCA [2017-2020])-Centre National de la Recherche Scientifique (CNRS) |
| Předmět: |
Polymers
[SDV]Life Sciences [q-bio] Nanotechnology Biology 010402 general chemistry 01 natural sciences Models Biological Antibodies 03 medical and health sciences Surface-Active Agents Native state [CHIM]Chemical Sciences Surface plasmon resonance Integral membrane protein ComputingMilieux_MISCELLANEOUS 030304 developmental biology 0303 health sciences Multidisciplinary Drug discovery Membrane Proteins Surface Plasmon Resonance Cross-Linking Reagents Microspheres 0104 chemical sciences Membrane Membrane protein Microscopy Fluorescence Proteome Physical Sciences Adsorption Snake Venoms |
| Zdroj: | University of Copenhagen Proceedings of the National Academy of Sciences of the United States of America Proceedings of the National Academy of Sciences of the United States of America, National Academy of Sciences, 2009, 106 (2), pp.405-410. ⟨10.1073/pnas.0807132106⟩ Proceedings of the National Academy of Sciences of the United States of America, 2009, 106 (2), pp.405-410. ⟨10.1073/pnas.0807132106⟩ Charvolin, D, Perez, J-B, Rouviere, F, Giusti, F, Bazzacco, P, Abdine, A, Rappaport, F, Martinez, K L & Popot, J-L 2009, ' The use of amphipols as universal molecular adapters to immobilize membrane proteins onto solid supports ', Proceedings of the National Academy of Science of the United States of America, vol. 106, no. 2, pp. 405-10 . |
| ISSN: | 0027-8424 1091-6490 |
| DOI: | 10.1073/pnas.0807132106⟩ |
| Popis: | Because of the importance of their physiological functions, cell membranes represent critical targets in biological research. Membrane proteins, which make up ≈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). |
| Databáze: | OpenAIRE |
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