Molecular Mechanisms of Class B GPCR Activation: Insights from Adrenomedullin Receptors.

Autor: Garelja ML; School of Biological Sciences, University of Auckland, Auckland, 1010, New Zealand., Au M; School of Biological Sciences, University of Auckland, Auckland, 1010, New Zealand.; Maurice Wilkins Centre for Molecular Biodiscovery, University of Auckland, Auckland, 1010, New Zealand., Brimble MA; School of Biological Sciences, University of Auckland, Auckland, 1010, New Zealand.; Maurice Wilkins Centre for Molecular Biodiscovery, University of Auckland, Auckland, 1010, New Zealand.; School of Chemical Sciences, University of Auckland, Auckland, 1010, New Zealand., Gingell JJ; School of Biological Sciences, University of Auckland, Auckland, 1010, New Zealand.; Maurice Wilkins Centre for Molecular Biodiscovery, University of Auckland, Auckland, 1010, New Zealand., Hendrikse ER; School of Biological Sciences, University of Auckland, Auckland, 1010, New Zealand., Lovell A; School of Biological Sciences, University of Auckland, Auckland, 1010, New Zealand., Prodan N; School of Biological Sciences, University of Auckland, Auckland, 1010, New Zealand., Sexton PM; Drug Discovery Biology and Department of Pharmacology, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria 3052, Australia., Siow A; School of Biological Sciences, University of Auckland, Auckland, 1010, New Zealand.; School of Chemical Sciences, University of Auckland, Auckland, 1010, New Zealand., Walker CS; School of Biological Sciences, University of Auckland, Auckland, 1010, New Zealand.; Maurice Wilkins Centre for Molecular Biodiscovery, University of Auckland, Auckland, 1010, New Zealand., Watkins HA; School of Biological Sciences, University of Auckland, Auckland, 1010, New Zealand.; Maurice Wilkins Centre for Molecular Biodiscovery, University of Auckland, Auckland, 1010, New Zealand., Williams GM; Maurice Wilkins Centre for Molecular Biodiscovery, University of Auckland, Auckland, 1010, New Zealand.; School of Chemical Sciences, University of Auckland, Auckland, 1010, New Zealand., Wootten D; Drug Discovery Biology and Department of Pharmacology, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria 3052, Australia., Yang SH; School of Biological Sciences, University of Auckland, Auckland, 1010, New Zealand.; Maurice Wilkins Centre for Molecular Biodiscovery, University of Auckland, Auckland, 1010, New Zealand., Harris PWR; School of Biological Sciences, University of Auckland, Auckland, 1010, New Zealand.; Maurice Wilkins Centre for Molecular Biodiscovery, University of Auckland, Auckland, 1010, New Zealand.; School of Chemical Sciences, University of Auckland, Auckland, 1010, New Zealand., Hay DL; School of Biological Sciences, University of Auckland, Auckland, 1010, New Zealand.; Maurice Wilkins Centre for Molecular Biodiscovery, University of Auckland, Auckland, 1010, New Zealand.
Jazyk: angličtina
Zdroj: ACS pharmacology & translational science [ACS Pharmacol Transl Sci] 2020 Feb 26; Vol. 3 (2), pp. 246-262. Date of Electronic Publication: 2020 Feb 26 (Print Publication: 2020).
DOI: 10.1021/acsptsci.9b00083
Abstrakt: Adrenomedullin (AM) is a 52 amino acid peptide that plays a regulatory role in the vasculature. Receptors for AM comprise the class B G protein-coupled receptor, the calcitonin-like receptor (CLR), in complex with one of three receptor activity-modifying proteins (RAMPs). The C-terminus of AM is involved in binding to the extracellular domain of the receptor, while the N-terminus is proposed to interact with the juxtamembranous portion of the receptor to activate signaling. There is currently limited information on the molecular determinants involved in AM signaling, thus we set out to define the importance of the AM N-terminus through five signaling pathways (cAMP production, ERK phosphorylation, CREB phosphorylation, Akt phosphorylation, and IP 1 production). We characterized the three CLR:RAMP complexes through the five pathways, finding that each had a distinct repertoire of intracellular signaling pathways that it is able to regulate. We then performed an alanine scan of AM from residues 15-31 and found that most residues could be substituted with only small effects on signaling, and that most substitutions affected signaling through all receptors and pathways in a similar manner. We identify F18, T20, L26, and I30 as being critical for AM function, while also identifying an analogue (AM 15-52 G19A) which has unique signaling properties relative to the unmodified AM. We interpret our findings in the context of new structural information, highlighting the complementary nature of structural biology and functional assays.
Competing Interests: The authors declare no competing financial interest.
(Copyright © 2020 American Chemical Society.)
Databáze: MEDLINE
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