title: Conformational Chaperones for Structural Studies of Membrane Proteins Using Antibody Phage Display with Nanodiscs.
abstract: A major challenge in membrane biophysics is to define the mechanistic linkages between a protein's conformational transitions and its function. We describe a novel approach to stabilize transient functional states of membrane proteins in native-like lipid environments allowing for their structural and biochemical characterization. This is accomplished by combining the power of antibody Fab-based phage display selection with the benefits of embedding membrane protein targets in lipid-filled nanodiscs. In addition to providing a stabilizing lipid environment, nanodiscs afford significant technical advantages over detergent-based formats. This enables the production of a rich pool of high-performance Fab binders that can be used as crystallization chaperones, as fiducial markers for single-particle cryoelectron microscopy, and as probes of different conformational states. Moreover, nanodisc-generated Fabs can be used to identify detergents that best mimic native membrane environments for use in biophysical studies.
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Chicago, 929 East 57th Street, Chicago, IL 60637, USA.
authors.forename: Pawel K
authors.initials: PK
authors.lastname: Dominik
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Chicago, 929 East 57th Street, Chicago, IL 60637, USA.
authors.forename: Marta T
authors.initials: MT
authors.lastname: Borowska
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Chicago, 929 East 57th Street, Chicago, IL 60637, USA.
authors.forename: Olivier
authors.initials: O
authors.lastname: Dalmas
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Chicago, 929 East 57th Street, Chicago, IL 60637, USA.
authors.forename: Sangwoo S
authors.initials: SS
authors.lastname: Kim
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Chicago, 929 East 57th Street, Chicago, IL 60637, USA.
authors.forename: Eduardo
authors.initials: E
authors.lastname: Perozo
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Chicago, 929 East 57th Street, Chicago, IL 60637, USA.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Keenan
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Chicago, 929 East 57th Street, Chicago, IL 60637, USA. Electronic address: koss@bsd.uchicago.edu.
authors.forename: Anthony A
authors.initials: AA
authors.lastname: Kossiakoff
mesh_terms: D061505:Cell Surface Display Techniques / Q000379:methods*; D003902:Detergents / Q000737:chemistry; D007128:Immunoglobulin Fragments / Q000737:chemistry*; D008051:Lipid Bilayers / Q000737:chemistry; D008565:Membrane Proteins / Q000737:chemistry*; D008958:Models, Molecular; D018832:Molecular Chaperones / Q000737:chemistry*; D049329:Nanostructures / Q000737:chemistry*; D019151:Peptide Library; D011487:Protein Conformation
chemical_list: D003902:Detergents; D007128:Immunoglobulin Fragments; D008051:Lipid Bilayers; D008565:Membrane Proteins; D018832:Molecular Chaperones; D019151:Peptide Library