title: Synthesis of an oligonucleotide-derivatized amphipol and its use to trap and immobilize membrane proteins.
abstract: Amphipols (APols) are specially designed amphipathic polymers that stabilize membrane proteins (MPs) in aqueous solutions in the absence of detergent. A8-35, a polyacrylate-based APol, has been grafted with an oligodeoxynucleotide (ODN). The synthesis, purification and properties of the resulting 'OligAPol' have been investigated. Grafting was performed by reacting an ODN carrying an amine-terminated arm with the carboxylates of A8-35. The use of OligAPol for trapping MPs and immobilizing them onto solid supports was tested using bacteriorhodopsin (BR) and the transmembrane domain of Escherichia coli outer membrane protein A (tOmpA) as model proteins. BR and OligAPol form water-soluble complexes in which BR remains in its native conformation. Hybridization of the ODN arm with a complementary ODN was not hindered by the assembly of OligAPol into particles, nor by its association with BR. BR/OligAPol and tOmpA/OligAPol complexes could be immobilized onto either magnetic beads or gold nanoparticles grafted with the complementary ODN, as shown by spectroscopic measurements, fluorescence microscopy and the binding of anti-BR and anti-tOmpA antibodies. OligAPols provide a novel, highly versatile approach to tagging MPs, without modifying them chemically nor genetically, for specific, reversible and targetable immobilization, e.g. for nanoscale applications.
authors.affiliation: UMR 7099, Centre National de la Recherche Scientifique/Université Paris-7, Institut de Biologie Physico-Chimique (FRC 550), 13 rue Pierre et Marie Curie, F-75005 Paris, France.
authors.forename: Christel
authors.initials: C
authors.lastname: Le Bon
authors.affiliation: Bio-Nanotechnology and Nanomedicine Laboratory, Department of Chemistry and Nano-Science Center, University of Copenhagen, Universitetsparken 5, DK-2100 Copenhagen, Denmark.
authors.forename: Eduardo Antonio
authors.initials: EA
authors.lastname: Della Pia
authors.affiliation: UMR 7099, Centre National de la Recherche Scientifique/Université Paris-7, Institut de Biologie Physico-Chimique (FRC 550), 13 rue Pierre et Marie Curie, F-75005 Paris, France.
authors.forename: Fabrice
authors.initials: F
authors.lastname: Giusti
authors.affiliation: Bio-Nanotechnology and Nanomedicine Laboratory, Department of Chemistry and Nano-Science Center, University of Copenhagen, Universitetsparken 5, DK-2100 Copenhagen, Denmark.
authors.forename: Noémie
authors.initials: N
authors.lastname: Lloret
authors.affiliation: UMR 7099, Centre National de la Recherche Scientifique/Université Paris-7, Institut de Biologie Physico-Chimique (FRC 550), 13 rue Pierre et Marie Curie, F-75005 Paris, France.
authors.forename: Manuela
authors.initials: M
authors.lastname: Zoonens
authors.affiliation: Bio-Nanotechnology and Nanomedicine Laboratory, Department of Chemistry and Nano-Science Center, University of Copenhagen, Universitetsparken 5, DK-2100 Copenhagen, Denmark.
authors.forename: Karen L
authors.initials: KL
authors.lastname: Martinez
authors.affiliation: UMR 7099, Centre National de la Recherche Scientifique/Université Paris-7, Institut de Biologie Physico-Chimique (FRC 550), 13 rue Pierre et Marie Curie, F-75005 Paris, France jean-luc.popot@ibpc.fr.
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Popot
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry; D001436:Bacteriorhodopsins / Q000737:chemistry; D006046:Gold; D055767:Immobilized Proteins / Q000737:chemistry; D008565:Membrane Proteins / Q000737:chemistry*; D053768:Metal Nanoparticles; D008863:Microspheres; D009693:Nucleic Acid Hybridization; D009838:Oligodeoxyribonucleotides / Q000737:chemistry*; D011108:Polymers / Q000138:chemical synthesis / Q000737:chemistry*; D011437:Propylamines / Q000138:chemical synthesis / Q000737:chemistry*
chemical_list: D001425:Bacterial Outer Membrane Proteins; D055767:Immobilized Proteins; D008565:Membrane Proteins; D009838:Oligodeoxyribonucleotides; D011108:Polymers; D011437:Propylamines; C499017:amphipol A8-35; C078167:OMPA outer membrane proteins; D001436:Bacteriorhodopsins; D006046:Gold
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