Conformational Chaperones for Structural Studies of Membrane Proteins Using Antibody Phage Display with Nanodiscs.

TitleConformational Chaperones for Structural Studies of Membrane Proteins Using Antibody Phage Display with Nanodiscs.
Publication TypeJournal Article
Year of Publication2016
AuthorsDominik, Pawel K., Borowska Marta T., Dalmas Olivier, Kim Sangwoo S., Perozo Eduardo, Keenan Robert J., and Kossiakoff Anthony A.
JournalStructure
Volume24
Issue2
Pagination300-9
Date Published2016 Feb 02
ISSN1878-4186
KeywordsCell Surface Display Techniques, Detergents, Immunoglobulin Fragments, Lipid Bilayers, Membrane Proteins, Models, Molecular, Molecular Chaperones, Nanostructures, Peptide Library, Protein Conformation
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.

DOI10.1016/j.str.2015.11.014
Alternate JournalStructure
PubMed ID26749445
PubMed Central IDPMC4740257
Grant ListR01GM086487 / GM / NIGMS NIH HHS / United States
R01 GM088406 / GM / NIGMS NIH HHS / United States
GM088406 / GM / NIGMS NIH HHS / United States
R01 GM086487 / GM / NIGMS NIH HHS / United States
U01 GM094588 / GM / NIGMS NIH HHS / United States
U54 GM087519 / GM / NIGMS NIH HHS / United States
R01 GM117372 / GM / NIGMS NIH HHS / United States

University of Toronto  UCSF  The University of Chicago  QB3  Chicago Biomedical Consortium