25-30 August 2019
Henry Ford Building
Europe/Berlin timezone

Insight into small molecule binding to the neonatal Fc receptor by X-ray crystallography and 100 kHz magic-angle-spinning NMR

28 Aug 2019, 13:45
10m
Max Kade Auditorium (Henry Ford Building)

Max Kade Auditorium

Henry Ford Building

Prize lecture Biological applications Prize Lectures

Speaker

Daniel Friedrich (Leibniz-Forschungsinstitut für Molekulare Pharmakologie, Berlin, Germany)

Description

Aiming at the design of an allosteric modulator of the neonatal Fc receptor (FcRn)–Immunoglobulin G (IgG) interaction, we developed a new methodology including NMR fragment screening, X-ray crystallography, and magic-angle-spinning (MAS) NMR at 100 kHz after sedimentation, exploiting very fast spinning of the nondeuterated soluble 42 kDa receptor construct to obtain resolved proton-detected 2D and 3D NMR spectra. FcRn plays a crucial role in regulation of IgG and serum albumin catabolism. It is a clinically validated drug target for the treatment of autoimmune diseases caused by pathogenic antibodies via the inhibition of its interaction with IgG. We herein present the discovery of a small molecule that binds into a conserved cavity of the heterodimeric, extracellular domain composed of an α-chain and β2-microglobulin (β2m) (FcRnECD, 373 residues). X-ray crystallography was used alongside NMR at 100 kHz MAS with sedimented soluble protein to explore possibilities for refining the compound as an allosteric modulator. Proton-detected MAS NMR experiments on fully protonated [13C,15N]-labeled FcRnECD yielded ligand-induced chemical-shift perturbations (CSPs) for residues in the binding pocket and allosteric changes close to the interface of the two receptor heterodimers present in the asymmetric unit as well as potentially in the albumin interaction site. X-ray structures with and without ligand suggest the need for an optimized compound to displace the α-chain with respect to β2m, both of which participate in the FcRnECD–IgG interaction site. Our investigation establishes a method to characterize structurally small molecule binding to nondeuterated large proteins by NMR, even in their glycosylated form, which may prove highly valuable for structure-based drug discovery campaigns.

Primary authors

Daniel Friedrich (Leibniz-Forschungsinstitut für Molekulare Pharmakologie, Berlin, Germany) Alex Macpherson (UCB Celltech, Slough, United Kingdom) Susanne Smith-Penzel (Laboratory of Physical Chemistry, ETH Zürich, Switzerland) Nicolas Basse (Sanofi, Strasbourg, France) Fabien Lecomte (UCB Celltech, Slough, United Kingdom) Hervé Deboves (Evotec, Milton, United Kingdom) Richard D. Taylor (UCB Celltech, Slough, United Kingdom) Tim Norman (UCB Celltech, Slough, United Kingdom) John Porter (Midatech Pharma Plc, Milton, United Kingdom) Lorna C. Waters (Institute of Structural and Chemical Biology, University of Leicester, United Kingdom) Marta Westwood (UCB Celltech, Slough, United Kingdom) Ben Cossins (UCB Celltech, Slough, United Kingdom) Katharine Cain (Vertex, Milton, United Kingdom) James White (UCB Celltech, Slough, United Kingdom) Robert Griffin (UCB Celltech, Slough, United Kingdom) Christine Prosser (UCB Celltech, Slough, United Kingdom) Sebastian Kelm (UCB Celltech, Slough, United Kingdom) Amy H. Sullivan (Beryllium Discovery, Bedford, Massachusetts, United States of America) David Fox III (Beryllium Discovery, Bedford, Massachusetts, United States of America) Mark D. Carr (Institute of Structural and Chemical Biology, University of Leicester, United Kingdom) Alistair Henry (UCB Celltech, Slough, United Kingdom) Richard Taylor (UCB Celltech, Slough, United Kingdom) Beat H. Meier (Laboratory of Physical Chemistry, ETH Zürich, Switzerland) Hartmut Oschkinat (Leibniz-Forschungsinstitut für Molekulare Pharmakologie, Berlin, Germany) Alastair D. Lawson (UCB Celltech, Slough, United Kingdom)

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