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- Title
Amphipol-Mediated Screening of Molecular Orthoses Specific for Membrane Protein Targets.
- Authors
Ferrandez, Yann; Dezi, Manuela; Bosco, Mickael; Urvoas, Agathe; Valerio-Lepiniec, Marie; Bon, Christel; Giusti, Fabrice; Broutin, Isabelle; Durand, Grégory; Polidori, Ange; Popot, Jean-Luc; Picard, Martin; Minard, Philippe
- 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.
- Subjects
DETERGENTS; MEMBRANE proteins; CRYSTALLIZATION; PROTEIN binding; PROTEIN stability; PROTEIN engineering
- Publication
Journal of Membrane Biology, 2014, Vol 247, Issue 9/10, p925
- ISSN
0022-2631
- Publication type
Article
- DOI
10.1007/s00232-014-9707-3