Nonionic homopolymeric amphipols: application to membrane protein folding, cell-free synthesis, and solution nuclear magnetic resonance
| Autor: | Grégory Durand, K. Shivaji Sharma, Laurent Catoire, Jean-Louis Banères, Emmanuelle Billon-Denis, Sophie Mary, Elodie Point, Francesca Zito, Christel Le Bon, Jean-Luc Popot, Bernard Pucci, Paola Bazzacco |
|---|---|
| Přispěvatelé: | Physico-chimie moléculaire des membranes biologiques (PCMMB), Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Institut des Biomolécules Max Mousseron [Pôle Chimie Balard] (IBMM), Ecole Nationale Supérieure de Chimie de Montpellier (ENSCM)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Unité de Chimie et Procédés (UCP), École Nationale Supérieure de Techniques Avancées (ENSTA Paris), Laboratoire de Chimie Bioorganique et des Systèmes Moléculaires Vectoriels (LCBOSMV), Avignon Université (AU), Laboratoire de biologie physico-chimique des protéines membranaires (LBPC-PM (UMR_7099)), Institut de biologie physico-chimique (IBPC (FR_550)), Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Montpellier (UM)-Ecole Nationale Supérieure de Chimie de Montpellier (ENSCM), Centre National de la Recherche Scientifique (CNRS)-Université Paris Diderot - Paris 7 (UPD7) |
| Rok vydání: | 2012 |
| Předmět: |
Halobacterium salinarum
Protein Folding Glycosylation Magnetic Resonance Spectroscopy Polymers [SDV]Life Sciences [q-bio] Chlamydomonas reinhardtii Buffers 010402 general chemistry 01 natural sciences Biochemistry 03 medical and health sciences chemistry.chemical_compound Nuclear magnetic resonance GTP-Binding Proteins Amide Native state Escherichia coli [CHIM]Chemical Sciences Receptors Ghrelin 030304 developmental biology Ions 0303 health sciences biology Cell-Free System Propylamines Membrane Proteins Bacteriorhodopsin biology.organism_classification Ghrelin 0104 chemical sciences Monomer Membrane protein chemistry Heteronuclear molecule Cytochromes b6 Bacteriorhodopsins biology.protein |
| Zdroj: | Biochemistry Biochemistry, American Chemical Society, 2012, 51 (7), pp.1416-1430. ⟨10.1021/bi201862v⟩ Biochemistry, American Chemical Society, 2012, ⟨10.1021/bi201862v⟩ |
| ISSN: | 1520-4995 0006-2960 |
| DOI: | 10.1021/bi201862v⟩ |
| Popis: | International audience; Nonionic amphipols (NAPols) synthesized by homotelomerization of an amphiphatic monomer are able to keep membrane proteins (MPs) stable and functional in the absence of detergent. Some of their biochemical and biophysical properties and applications have been examined, with particular attention being paid to their complementarity with the classical polyacrylate-based amphipol A8-35. Bacter-iorhodopsin (BR) from Halobacterium salinarum and the cytochrome b 6 f complex from Chlamydomonas reinhardtii were found to be in their native state and highly stable following complexation with NAPols. NAPol-trapped BR was shown to undergo its complete photocycle. Because of the pH insensitivity of NAPols, solution nuclear magnetic resonance (NMR) two-dimensional 1 H− 15 N heteronuclear single-quantum coherence spectra of NAPol-trapped outer MP X from Escherichia coli (OmpX) could be recorded at pH 6.8. They present a resolution similar to that of the spectra of OmpX/A8-35 complexes recorded at pH 8.0 and give access to signals from solvent-exposed rapidy exchanging amide protons. Like A8-35, NAPols can be used to fold MPs to their native state as demonstrated here with BR and with the ghrelin G protein-coupled receptor GHS-R1a, thus extending the range of accessible folding conditions. Following NAPol-assisted folding, GHS-R1a bound four of its specific ligands, recruited arrestin-2, and activated binding of GTPγS by the G αq protein. Finally, cell-free synthesis of MPs, which is inhibited by A8-35 and sulfonated amphipols, was found to be very efficient in the presence of NAPols. These results open broad new perspectives on the use of amphipols for MP studies. S |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|