scholarly journals A distinctive ligand recognition mechanism by the human vasoactive intestinal polypeptide receptor 2

2021 ◽  
Author(s):  
Yingna Xu ◽  
Wenbo Feng ◽  
Qingtong Zhou ◽  
Anyi Liang ◽  
Jie Li ◽  
...  
Keyword(s):  
G Protein ◽  
Vasoactive Intestinal Polypeptide ◽  
Drug Targets ◽  
Receptor Activation ◽  
Dimensional Structure ◽  
Recognition Mechanism ◽  
Additional Information ◽  
Stimulatory G Protein ◽  
Α Helix ◽  
Intestinal Polypeptide

Activated by physiologically important peptide hormones, class B1 G protein-coupled receptors (GPCRs) modulate key physiological functions and serve as valuable drug targets for many diseases. Among them, vasoactive intestinal polypeptide receptor 2 (VIP2R) is the last member whose full-length 3-dimensional structure has yet to be determined. VIP2R, expressed in the central and peripheral nervous systems and involved in a number of pathophysiological conditions, is implicated in pulmonary arterial hypertension, autoimmune and psychiatric disorders. Here, we report the cryo-electron microscopy structure of the human VIP2R bound to its endogenous ligand PACAP27 and the stimulatory G protein. Different from all reported peptide-bound class B1 GPCR structures, the N-terminal α-helix of VIP2R adopts a unique conformation that deeply inserts into a cleft between PACAP27 and the extracellular loop 1, thereby stabilizing the peptide-receptor interface. Its truncation or extension significantly decreased VIP2R-mediated cAMP accumulation. Our results provide additional information on peptide recognition and receptor activation among class B1 GPCRs and may facilitate the design of better therapeutics.

Constitutive Formation of Oligomeric Complexes between Family B G Protein-Coupled Vasoactive Intestinal Polypeptide and Secretin Receptors

2005 ◽  
Vol 69 (1) ◽  
pp. 363-373 ◽  
Author(s):  
Kaleeckal G. Harikumar ◽  
Maria M. Morfis ◽  
Cayle S. Lisenbee ◽  
Patrick M. Sexton ◽  
Laurence J. Miller
Keyword(s):  
G Protein ◽  
Vasoactive Intestinal Polypeptide ◽  
G Protein Coupled ◽  
Intestinal Polypeptide

scholarly journals SNAP-tagged nanobodies enable reversible optical control of a G protein-coupled receptor via a remotely tethered photoswitchable ligand

2018 ◽  
Author(s):  
Helen Farrants ◽  
Amanda Acosta Ruiz ◽  
Vanessa A. Gutzeit ◽  
Dirk Trauner ◽  
Kai Johnsson ◽  
...  
Keyword(s):  
G Protein ◽  
Drug Targets ◽  
Metabotropic Glutamate Receptor ◽  
Receptor Activation ◽  
Metabotropic Glutamate ◽  
Extracellular Signals ◽  
Physiological Processes ◽  
Wide Range ◽  
G Protein Coupled

AbstractG protein-coupled receptors (GPCRs) mediate the transduction of extracellular signals into complex intracellular responses. Despite their ubiquitous roles in physiological processes and as drug targets for a wide range of disorders, the precise mechanisms of GPCR function at the molecular, cellular, and systems levels remain partially understood. In order to dissect the function of individual receptors subtypes with high spatiotemporal precision, various optogenetic and photopharmacological approaches have been reported that use the power of light for receptor activation and deactivation. Here, we introduce a novel and, to date, most remote way of applying photoswitchable orthogonally remotely-tethered ligands (PORTLs) by using a SNAP-tag fused nanobody. Our nanobody-photoswitch conjugates (NPCs) can be used to target a GFP-fused metabotropic glutamate receptor by either gene-free application of purified complexes or co-expression of genetically encoded nanobodies to yield robust, reversible control of agonist binding and subsequent downstream activation. By harboring and combining the selectivity and flexibility of both nanobodies and self-labelling enzymes, we set the stage for targeting endogenous receptors in vivo.

scholarly journals Multiplex GPCR Assay in Reverse Transfection Cell Microarrays

2004 ◽  
Vol 9 (3) ◽  
pp. 196-207 ◽  
Author(s):  
Yuji M. Mishina ◽  
Christopher J. Wilson ◽  
Linda Bruett ◽  
Jesse J. Smith ◽  
Chatanika Stoop-Myer ◽  
...  
Keyword(s):  
G Protein ◽  
Drug Targets ◽  
Receptor Activation ◽  
Calcium Flux ◽  
Orphan Receptors ◽  
Ligand Selectivity ◽  
Ligand Interactions ◽  
Cell Microarrays ◽  
Reverse Transfection ◽  
Targeted Drug Discovery

G protein-coupled receptors (GPCRs) are a superfamily of proteins that include some of the most important drug targets in the pharmaceutical industry. Despite the success of this group of drugs, there remains a need to identify GPCR-targeted drugs with greater selectivity, to develop screening assays for validated targets, and to identify ligands for orphan receptors. To address these challenges, the authors have created a multiplexed GPCR assay that measures greater than 3000 receptor: ligand interactions in a single microplate. The multiplexed assay is generated by combining reverse transfection in a 96-well plate format with a calcium flux readout. This assay quantitatively measures receptor activation and inhibition and permits the determination of compound potency and selectivity for entire families of GPCRs in parallel. To expand the number of GPCR targets that may be screened in this system, receptors are cotransfected with plasmids encoding a promiscuous G protein, permitting the analysis of receptors that do not normally mobilize intracellular calcium upon activation. The authors demonstrate the utility of reverse transfection cell microarrays to GPCR-targeted drug discovery with examples of ligand selectivity screening against a panel of GPCRs as well as dose-dependent titrations of selected agonists and antagonists.

Structural, signalling and regulatory properties of the group I metabotropic glutamate receptors: prototypic family C G-protein-coupled receptors

2001 ◽  
Vol 359 (3) ◽  
pp. 465-484 ◽  
Author(s):  
Emmanuel HERMANS ◽  
R. A. John CHALLISS
Keyword(s):  
G Protein ◽  
Drug Targets ◽  
Metabotropic Glutamate Receptors ◽  
Gabab Receptor ◽  
Receptor Activation ◽  
Metabotropic Glutamate ◽  
Calcium Sensing ◽  
Group I ◽  
G Protein Coupled ◽  
The Brain

In 1991 a new type of G-protein-coupled receptor (GPCR) was cloned, the type 1a metabotropic glutamate (mGlu) receptor, which, despite possessing the defining seven-transmembrane topology of the GPCR superfamily, bore little resemblance to the growing number of other cloned GPCRs. Subsequent studies have shown that there are eight mammalian mGlu receptors that, together with the calcium-sensing receptor, the GABAB receptor (where GABA is γ-aminobutyric acid) and a subset of pheromone, olfactory and taste receptors, make up GPCR family C. Currently available data suggest that family C GPCRs share a number of structural, biochemical and regulatory characteristics, which differ markedly from those of the other GPCR families, most notably the rhodopsin/family A GPCRs that have been most widely studied to date. This review will focus on the group I mGlu receptors (mGlu1 and mGlu5). This subgroup of receptors is widely and differentially expressed in neuronal and glial cells within the brain, and receptor activation has been implicated in the control of an array of key signalling events, including roles in the adaptative changes needed for long-term depression or potentiation of neuronal synaptic connectivity. In addition to playing critical physiological roles within the brain, the mGlu receptors are also currently the focus of considerable attention because of their potential as drug targets for the treatment of a variety of neurological and psychiatric disorders.

scholarly journals Evolution of Bovine Respiratory Syncytial Virus

2000 ◽  
Vol 74 (22) ◽  
pp. 10714-10728 ◽  
Author(s):  
Jean-François Valarcher ◽  
François Schelcher ◽  
Hervé Bourhy
Keyword(s):  
Respiratory Syncytial Virus ◽  
G Protein ◽  
Three Dimensional ◽  
Bovine Respiratory Syncytial Virus ◽  
Amino Acid Sequences ◽  
Dimensional Structure ◽  
Protein Fusion ◽  
N Protein ◽  
Syncytial Virus ◽  
Α Helix

ABSTRACT Until now, the analysis of the genetic diversity of bovine respiratory syncytial virus (BRSV) has been based on small numbers of field isolates. In this report, we determined the nucleotide and deduced amino acid sequences of regions of the nucleoprotein (N protein), fusion protein (F protein), and glycoprotein (G protein) of 54 European and North American isolates and compared them with the sequences of 33 isolates of BRSV obtained from the databases, together with those of 2 human respiratory syncytial viruses and 1 ovine respiratory syncytial virus. A clustering of BRSV sequences according to geographical origin was observed. We also set out to show that a continuous evolution of the sequences of the N, G, and F proteins of BRSV has been occurring in isolates since 1967 in countries where vaccination was widely used. The exertion of a strong positive selective pressure on the mucin-like region of the G protein and on particular sites of the N and F proteins is also demonstrated. Furthermore, mutations which are located in the conserved central hydrophobic part of the ectodomain of the G protein and which result in the loss of four Cys residues and in the suppression of two disulfide bridges and an α helix critical to the three-dimensional structure of the G protein have been detected in some recent French BRSV isolates. This conserved central region, which is immunodominant in BRSV G protein, thus has been modified in recent isolates. This work demonstrates that the evolution of BRSV should be taken into account in the rational development of future vaccines.

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