20
IRUS Total
Downloads

Modulation of PTH1R signaling by an ECD binding antibody results in inhibition of beta-arrestin 2 coupling

Title: Modulation of PTH1R signaling by an ECD binding antibody results in inhibition of beta-arrestin 2 coupling
Authors: Sarkar, K
Joedicke, L
Westwood, M
Burnley, R
Wright, M
McMillan, D
Byrne, B
Item Type: Journal Article
Abstract: Parathyroid hormone receptor 1 (PTH1R) belongs to the secretin class of G protein coupled receptors (GPCRs) and natively binds parathyroid hormone (PTH) and parathyroid hormone related peptide (PTHrP). Ligand binding to PTH1R involves binding to the large extracellular domain (ECD) and the orthosteric pocket, inducing conformational changes in the transmembrane domain and receptor activation. PTH1R regulates bone metabolism, signaling mainly through Gs and Gq/11 G-proteins. Here, we used phage display to generate PTH1R ECD-specific antibodies with the aim of modulating receptor functionality. We identified ECD-scFvhFc, which exhibited high affinity binding to both the isolated ECD and to the full-length receptor in styrene-maleic acid (SMA) lipid particles. Epitope mapping using hydrogen-deuterium exchange mass spectrometry (HDX-MS) indicates that the α1 helix of the ECD is ECD-scFvhFc’s epitope which may partially overlap with the known PTH (1–34) binding site. However, PTH (1–34)-mediated Gs activation is Undisturbed by ECD-scFvhFc binding. In contrast, ECD-scFvhFc potently inhibits β-arrestin-2 recruitment after PTH (1–34)-driven receptor activation and thus represents the first monoclonal antibody to selectively inhibit distinct PTH1R signaling pathways. Given the complexity of PTH1R signaling and the emerging importance of biased GPCR activation in drug development, ECD-scFvhFc could be a valuable tool to study PTH1R signaling bias.
Issue Date: 8-Oct-2019
Date of Acceptance: 24-Sep-2019
URI: http://hdl.handle.net/10044/1/76860
DOI: 10.1038/s41598-019-51016-z
ISSN: 2045-2322
Publisher: Nature Publishing Group
Journal / Book Title: Scientific Reports
Volume: 9
Copyright Statement: © The Author(s) 2019. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. Te images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
Keywords: Science & Technology
Multidisciplinary Sciences
Science & Technology - Other Topics
PARATHYROID-HORMONE RECEPTOR
C ACTIVATION DOMAIN
MEMBRANE-PROTEINS
INTERMITTENT TREATMENT
PEPTIDE
DIFFERENTIATION
FRAGMENT
AGONIST
INTACT
Science & Technology
Multidisciplinary Sciences
Science & Technology - Other Topics
PARATHYROID-HORMONE RECEPTOR
C ACTIVATION DOMAIN
MEMBRANE-PROTEINS
INTERMITTENT TREATMENT
PEPTIDE
DIFFERENTIATION
FRAGMENT
AGONIST
INTACT
Publication Status: Published
Article Number: ARTN 14432
Online Publication Date: 2019-10-08
Appears in Collections:Faculty of Natural Sciences