G Protein Coupling and Ligand Selectivity of the D2L and D3 Dopamine Receptors

2008 ◽  
Vol 325 (1) ◽  
pp. 319-330 ◽  
Author(s):  
J. Robert Lane ◽  
Ben Powney ◽  
Alan Wise ◽  
Stephen Rees ◽  
Graeme Milligan
Keyword(s):  
Dopamine Receptors ◽  
G Protein ◽  
G Protein Coupling ◽  
Protein Coupling ◽  
Ligand Selectivity

scholarly journals Ligand recognition, unconventional activation, and G protein coupling of the prostaglandin E2 receptor EP2 subtype

2021 ◽  
Vol 7 (14) ◽  
pp. eabf1268
Author(s):  
Changxiu Qu ◽  
Chunyou Mao ◽  
Peng Xiao ◽  
Qingya Shen ◽  
Ya-Ni Zhong ◽  
...  
Keyword(s):  
G Protein ◽  
Rational Design ◽  
Receptor Activation ◽  
Prostaglandin E ◽  
Toggle Switch ◽  
G Protein Coupling ◽  
Protein Coupling ◽  
Ligand Selectivity ◽  
Activation Mechanisms ◽  
Prostaglandin E2 Receptor

Selective modulation of the heterotrimeric G protein α S subunit–coupled prostaglandin E2 (PGE2) receptor EP2 subtype is a promising therapeutic strategy for osteoporosis, ocular hypertension, neurodegenerative diseases, and cardiovascular disorders. Here, we report the cryo–electron microscopy structure of the EP2-Gs complex with its endogenous agonist PGE2 and two synthesized agonists, taprenepag and evatanepag (CP-533536). These structures revealed distinct features of EP2 within the EP receptor family in terms of its unconventional receptor activation and G protein coupling mechanisms, including activation in the absence of a typical W6.48 “toggle switch” and coupling to Gs via helix 8. Moreover, inspection of the agonist-bound EP2 structures uncovered key motifs governing ligand selectivity. Our study provides important knowledge for agonist recognition and activation mechanisms of EP2 and will facilitate the rational design of drugs targeting the PGE2 signaling system.

scholarly journals Coupling of D2 dopamine receptors to G-proteins in solubilized preparations of bovine caudate nucleus

1992 ◽  
Vol 281 (2) ◽  
pp. 369-375 ◽  
Author(s):  
J A Chazot ◽  
P G Strange
Keyword(s):  
Ionic Strength ◽  
Caudate Nucleus ◽  
Dopamine Receptors ◽  
G Protein ◽  
G Proteins ◽  
Gel Filtration ◽  
D2 Dopamine Receptors ◽  
G Protein Coupling ◽  
Protein Coupling ◽  
Primary Determinant

1. The coupling of D2 dopamine receptors and G-proteins has been examined in cholate-solubilized preparations of bovine caudate nucleus. 2. No receptor-G-protein coupling could be detected in solubilized preparations obtained in 0.3% cholate, but if this preparation is diluted 5-fold, coupling is re-established. 3. The dilution process was examined, and it was shown that the change in ionic strength was an important factor in modulating the observed receptor-G-protein interaction. 4. Ionic strength was shown, however, not to be the primary determinant of receptor-G-protein coupling. This is likely to be the formation, upon dilution of the preparation, of vesicles in which receptor and G-protein reassociate. 5. The formation of vesicles upon dilution was examined by a variety of techniques, including thermal-stability studies, gel filtration, centrifugation and electron microscopy.

scholarly journals Biochemical characterization of G protein coupling to calcitonin gene–related peptide and adrenomedullin receptors using a native PAGE assay

2020 ◽  
Vol 295 (28) ◽  
pp. 9736-9751 ◽  
Author(s):  
Amanda M. Roehrkasse ◽  
Margaret L. Warner ◽  
Jason M. Booe ◽  
Augen A. Pioszak
Keyword(s):  
G Protein ◽  
Mammalian Cells ◽  
Calcitonin Gene Related Peptide ◽  
Peptide Ligand ◽  
Native Page ◽  
G Protein Coupling ◽  
Protein Coupling ◽  
Ligand Selectivity ◽  
Related Peptide ◽  
Calcitonin Gene

Calcitonin gene-related peptide (CGRP), adrenomedullin (AM), and adrenomedullin 2/intermedin (AM2/IMD) have overlapping and unique functions in the nervous and circulatory systems including vasodilation, cardioprotection, and pain transmission. Their actions are mediated by the class B calcitonin-like G protein–coupled receptor (CLR), which heterodimerizes with three receptor activity–modifying proteins (RAMP1–3) that determine its peptide ligand selectivity. How the three agonists and RAMPs modulate CLR binding to transducer proteins remains poorly understood. Here, we biochemically characterized agonist-promoted G protein coupling to each CLR·RAMP complex. We adapted a native PAGE method to assess the formation and thermostabilities of detergent-solubilized fluorescent protein–tagged CLR·RAMP complexes expressed in mammalian cells. Addition of agonist and the purified Gs protein surrogate mini-Gs (mGs) yielded a mobility-shifted agonist·CLR·RAMP·mGs quaternary complex gel band that was sensitive to antagonists. Measuring the apparent affinities of the agonists for the mGs-coupled receptors and of mGs for the agonist-occupied receptors revealed that both ligand and RAMP control mGs coupling and defined how agonist engagement of the CLR extracellular and transmembrane domains affects transducer recruitment. Using mini-Gsq and -Gsi chimeras, we observed a coupling rank order of mGs > mGsq > mGsi for each receptor. Last, we demonstrated the physiological relevance of the native gel assays by showing that they can predict the cAMP-signaling potencies of AM and AM2/IMD chimeras. These results highlight the power of the native PAGE assay for membrane protein biochemistry and provide a biochemical foundation for understanding the molecular basis of shared and distinct signaling properties of CGRP, AM, and AM2/IMD.

Functional Domains of the Mouse β3-Adrenoceptor Associated with Differential G Protein Coupling

2005 ◽  
Vol 315 (3) ◽  
pp. 1354-1361 ◽  
Author(s):  
Masaaki Sato ◽  
Dana S. Hutchinson ◽  
Tore Bengtsson ◽  
Anders Floren ◽  
Ülo Langel ◽  
...  
Keyword(s):  
G Protein ◽  
Functional Domains ◽  
G Protein Coupling ◽  
Protein Coupling

scholarly journals Dynamics of receptor/G protein coupling in living cells

2005 ◽  
Vol 24 (23) ◽  
pp. 4106-4114 ◽  
Author(s):  
Peter Hein ◽  
Monika Frank ◽  
Carsten Hoffmann ◽  
Martin J Lohse ◽  
Moritz Bünemann
Keyword(s):  
G Protein ◽  
Living Cells ◽  
G Protein Coupling ◽  
Protein Coupling

Mutant M1 muscarinic receptors with increased agonist affinity and G protein coupling

Life Sciences ◽  
1999 ◽  
Vol 64 (6-7) ◽  
pp. 563
Author(s):  
W.S. Messer ◽  
X.-P. Huang ◽  
P.I. Nagy ◽  
F.E. Williams ◽  
S.M. Peseckis
Keyword(s):  
G Protein ◽  
Muscarinic Receptors ◽  
G Protein Coupling ◽  
Protein Coupling ◽  
Agonist Affinity ◽  
M1 Muscarinic ◽  
M1 Muscarinic Receptors
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