title: Amphipol-mediated screening of molecular orthoses specific for membrane protein targets.
abstract: Specific, tight-binding protein partners are valuable helpers to facilitate membrane protein (MP) crystallization, because they can i) stabilize the protein, ii) reduce its conformational heterogeneity, and iii) increase the polar surface from which well-ordered crystals can grow. The design and production of a new family of synthetic scaffolds (dubbed αReps, for "artificial alpha repeat protein") have been recently described. The stabilization and immobilization of MPs in a functional state are an absolute prerequisite for the screening of binders that recognize specifically their native conformation. We present here a general procedure for the selection of αReps specific of any MP. It relies on the use of biotinylated amphipols, which act as a universal "Velcro" to stabilize, and immobilize MP targets onto streptavidin-coated solid supports, thus doing away with the need to tag the protein itself.
authors.affiliation: Laboratoire de Modélisation et Ingénierie des Protéines, IBBMC UMR 8619, CNRS/Université Paris Sud, 91405, Orsay, France.
authors.forename: Yann
authors.initials: Y
authors.lastname: Ferrandez
authors.forename: Manuela
authors.initials: M
authors.lastname: Dezi
authors.forename: Mickael
authors.initials: M
authors.lastname: Bosco
authors.forename: Agathe
authors.initials: A
authors.lastname: Urvoas
authors.forename: Marie
authors.initials: M
authors.lastname: Valerio-Lepiniec
authors.forename: Christel
authors.initials: C
authors.lastname: Le Bon
authors.forename: Fabrice
authors.initials: F
authors.lastname: Giusti
authors.forename: Isabelle
authors.initials: I
authors.lastname: Broutin
authors.forename: Grégory
authors.initials: G
authors.lastname: Durand
authors.forename: Ange
authors.initials: A
authors.lastname: Polidori
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Popot
authors.forename: Martin
authors.initials: M
authors.lastname: Picard
authors.forename: Philippe
authors.initials: P
authors.lastname: Minard
mesh_terms: D002462:Cell Membrane / Q000737:chemistry*; D057927:Hydrophobic and Hydrophilic Interactions; D008565:Membrane Proteins / Q000032:analysis / Q000737:chemistry*; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry*; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D021381:Protein Transport; D012995:Solubility; D013501:Surface-Active Agents / Q000737:chemistry*; D054457:Tissue Scaffolds / Q000737:chemistry
chemical_list: D008565:Membrane Proteins; D019151:Peptide Library; D010455:Peptides; D013501:Surface-Active Agents
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