scholarly journals Identification and mechanism of G protein-biased ligands for chemokine receptor CCR1

2021 ◽  
Author(s):  
Zhehua Shao ◽  
Qingya Shen ◽  
Bingpeng Yao ◽  
Chunyou Mao ◽  
Li-Nan Chen ◽  
...  
Keyword(s):  
G Protein ◽  
Chemokine Receptor ◽  
Binding Pocket ◽  
Structural Basis ◽  
Free Form ◽  
Downstream Signaling ◽  
Receptor Complexes ◽  
Ligand Free ◽  
Biased Signaling ◽  
G Protein Coupled

AbstractBiased signaling of G protein-coupled receptors describes an ability of different ligands that preferentially activate an alternative downstream signaling pathway. In this work, we identified and characterized different N-terminal truncations of endogenous chemokine CCL15 as balanced or biased agonists targeting CCR1, and presented three cryogenic-electron microscopy structures of the CCR1–Gi complex in the ligand-free form or bound to different CCL15 truncations with a resolution of 2.6–2.9 Å, illustrating the structural basis of natural biased signaling that initiates an inflammation response. Complemented with pharmacological and computational studies, these structures revealed it was the conformational change of Tyr291 (Y2917.43) in CCR1 that triggered its polar network rearrangement in the orthosteric binding pocket and allosterically regulated the activation of β-arrestin signaling. Our structure of CCL15-bound CCR1 also exhibited a critical site for ligand binding distinct from many other chemokine–receptor complexes, providing new insights into the mode of chemokine recognition.

scholarly journals Angiotensin and biased analogs induce structurally distinct active conformations within a GPCR

Science ◽  
2020 ◽  
Vol 367 (6480) ◽  
pp. 888-892 ◽  
Author(s):  
Laura M. Wingler ◽  
Meredith A. Skiba ◽  
Conor McMahon ◽  
Dean P. Staus ◽  
Alissa L. W. Kleinhenz ◽  
...  
Keyword(s):  
Angiotensin Ii ◽  
Chemokine Receptors ◽  
Binding Pocket ◽  
Protein Signaling ◽  
Downstream Signaling ◽  
The Family ◽  
Biased Signaling ◽  
Sodium Binding ◽  
G Protein Coupled

Biased agonists of G protein–coupled receptors (GPCRs) preferentially activate a subset of downstream signaling pathways. In this work, we present crystal structures of angiotensin II type 1 receptor (AT1R) (2.7 to 2.9 angstroms) bound to three ligands with divergent bias profiles: the balanced endogenous agonist angiotensin II (AngII) and two strongly β-arrestin–biased analogs. Compared with other ligands, AngII promotes more-substantial rearrangements not only at the bottom of the ligand-binding pocket but also in a key polar network in the receptor core, which forms a sodium-binding site in most GPCRs. Divergences from the family consensus in this region, which appears to act as a biased signaling switch, may predispose the AT1R and certain other GPCRs (such as chemokine receptors) to adopt conformations that are capable of activating β-arrestin but not heterotrimeric Gq protein signaling.

scholarly journals Engineering of Nanobodies Recognizing the Human Chemokine Receptor CCR7

2019 ◽  
Vol 20 (10) ◽  
pp. 2597 ◽  
Author(s):  
Barbara D. Jakobs ◽  
Lisa Spannagel ◽  
Vladimir Purvanov ◽  
Edith Uetz-von Allmen ◽  
Christoph Matti ◽  
...  
Keyword(s):  
T Cells ◽  
G Protein ◽  
Chemokine Receptor ◽  
Bimolecular Fluorescence Complementation ◽  
Downstream Signaling ◽  
Adaptive Immune ◽  
Health And Disease ◽  
Β2 Adrenergic Receptor ◽  
G Protein Coupled ◽  
Complementarity Determining Region

The chemokine receptor CCR7 plays a pivotal role in health and disease. In particular, CCR7 controls homing of antigen-bearing dendritic cells and T cells to lymph nodes, where adaptive immune responses are initiated. However, CCR7 also guides T cells to inflamed synovium and thereby contributes to rheumatoid arthritis and promotes cancer cell migration and metastasis formation. Nanobodies have recently emerged as versatile tools to study G-protein-coupled receptor functions and are being tested in diagnostics and therapeutics. In this study, we designed a strategy to engineer novel nanobodies recognizing human CCR7. We generated a nanobody library based on a solved crystal structure of the nanobody Nb80 recognizing the β2-adrenergic receptor (β2AR) and by specifically randomizing two segments within complementarity determining region 1 (CDR1) and CDR3 of Nb80 known to interact with β2AR. We fused the nanobody library to one half of split-YFP in order to identify individual nanobody clones interacting with CCR7 fused to the other half of split-YFP using bimolecular fluorescence complementation. We present three novel nanobodies, termed Nb1, Nb5, and Nb38, that recognize human CCR7 without interfering with G-protein-coupling and downstream signaling. Moreover, we were able to follow CCR7 trafficking upon CCL19 triggering using Nb1, Nb5, and Nb38.

scholarly journals Molecular mechanism of biased signaling in a prototypical G protein–coupled receptor

Science ◽  
2020 ◽  
Vol 367 (6480) ◽  
pp. 881-887 ◽  
Author(s):  
Carl-Mikael Suomivuori ◽  
Naomi R. Latorraca ◽  
Laura M. Wingler ◽  
Stephan Eismann ◽  
Matthew C. King ◽  
...  
Keyword(s):  
G Protein ◽  
Protein A ◽  
Binding Pocket ◽  
Great Promise ◽  
G Protein Coupled Receptor ◽  
Structural Mechanism ◽  
Biased Signaling ◽  
Dynamics Simulations ◽  
Effective Drugs ◽  
G Protein Coupled

Biased signaling, in which different ligands that bind to the same G protein–coupled receptor preferentially trigger distinct signaling pathways, holds great promise for the design of safer and more effective drugs. Its structural mechanism remains unclear, however, hampering efforts to design drugs with desired signaling profiles. Here, we use extensive atomic-level molecular dynamics simulations to determine how arrestin bias and G protein bias arise at the angiotensin II type 1 receptor. The receptor adopts two major signaling conformations, one of which couples almost exclusively to arrestin, whereas the other also couples effectively to a G protein. A long-range allosteric network allows ligands in the extracellular binding pocket to favor either of the two intracellular conformations. Guided by this computationally determined mechanism, we designed ligands with desired signaling profiles.

scholarly journals Endogenous agonist–bound S1PR3 structure reveals determinants of G protein–subtype bias

2021 ◽  
Vol 7 (24) ◽  
pp. eabf5325
Author(s):  
Shintaro Maeda ◽  
Yuki Shiimura ◽  
Hidetsugu Asada ◽  
Kunio Hirata ◽  
Fangjia Luo ◽  
...  
Keyword(s):  
G Protein ◽  
G Protein Coupled Receptors ◽  
Structural Basis ◽  
Active Conformation ◽  
Vascular Integrity ◽  
Biased Signaling ◽  
Sphingosine 1 Phosphate ◽  
G Protein Coupled ◽  
Alkyl Tail ◽  
Insight Into

Sphingosine-1-phosphate (S1P) regulates numerous important physiological functions, including immune response and vascular integrity, via its cognate receptors (S1PR1 to S1PR5); however, it remains unclear how S1P activates S1PRs upon binding. Here, we determined the crystal structure of the active human S1PR3 in complex with its natural agonist S1P at 3.2-Å resolution. S1P exhibits an unbent conformation in the long tunnel, which penetrates through the receptor obliquely. Compared with the inactive S1PR1 structure, four residues surrounding the alkyl tail of S1P (the “quartet core”) exhibit orchestrating rotamer changes that accommodate the moiety, thereby inducing an active conformation. In addition, we reveal that the quartet core determines G protein selectivity of S1PR3. These results offer insight into the structural basis of activation and biased signaling in G protein–coupled receptors and will help the design of biased ligands for optimized therapeutics.

scholarly journals The G Protein-Coupled Receptor Kinases (GRKs) in Chemokine Receptor-Mediated Immune Cell Migration: From Molecular Cues to Physiopathology

Cells ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 75
Author(s):  
Marta Laganà ◽  
Géraldine Schlecht-Louf ◽  
Françoise Bachelerie
Keyword(s):  
G Protein ◽  
Chemokine Receptor ◽  
Immune Cell ◽  
Neutrophil Migration ◽  
G Protein Coupled Receptor ◽  
Receptor Kinases ◽  
Immune Cell Migration ◽  
And Migration ◽  
G Protein Coupled ◽  
Gpcr Desensitization

Although G protein-coupled receptor kinases (GRKs) have long been known to regulate G protein-coupled receptor (GPCR) desensitization, their more recently characterized functions as scaffolds and signalling adapters underscore that this small family of proteins governs a larger array of physiological functions than originally suspected. This review explores how GRKs contribute to the complex signalling networks involved in the migration of immune cells along chemokine gradients sensed by cell surface GPCRs. We outline emerging evidence indicating that the coordinated docking of several GRKs on an active chemokine receptor determines a specific receptor phosphorylation barcode that will translate into distinct signalling and migration outcomes. The guidance cues for neutrophil migration are emphasized based on several alterations affecting GRKs or GPCRs reported to be involved in pathological conditions.

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