Nucleotide-Mediated Mucin Secretion from Differentiated Human Bronchial Epithelial Cells

Kemp, P.A.; Sugar, Rosemary; Jackson, A D

doi:10.1165/rcmb.2003-0211oc

Cited by 82 publications

(89 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Kemp et al [27] found it to be inactive at a concentration of 10 μM on a number of different receptors. These included the following human GPCRs: the serotonin receptors 5-HT 1A , 5-HT 2A , 5-HT 2B , 5-HT 2C , 5-HT 6 , and 5-HT 7 , the adrenergic receptors α 1 , α 2A , α 2B , α 2C , β 1 , and β 2 , the cannabinoid receptor CB 1 , the dopamine receptors D 1 , D 2 Lh, D 3 , and D 4 , the muscarinic acetylcholine receptors M 1 , M 2 , M 3 , M 4 , and M 5 , the histamine receptors H 1 and H 2 , the tachykinin receptors NK 1 and NK 2 , the opioid receptors δ, κ, and μ, GABA B , as well as the rat P2Y 1 receptor.…”

Section: Pharmacological Evaluation Of Ar-c118925 (3) and Its Derivatmentioning

confidence: 99%

“…The panel further included the following human ion channels: 5-HT 3 , L-type and N-type calcium and ATP-sensitive potassium channels. They further assessed the human P2Y 2 receptor for antagonism in a sandwich enzyme-linked lectin assay measuring mucin secretion, where they determined an IC 50 value of approximately 1 μM upon receptor stimulation with 100 μM ATPγS [27].…”

Section: Pharmacological Evaluation Of Ar-c118925 (3) and Its Derivatmentioning

confidence: 99%

“…One of the most selective antagonists described to date is the thiouracil derivative 5- Fig. 1) [26,27], which has been developed by structural modifications of the physiological agonist UTP [28]. Replacement of the ribose triphosphate moiety and derivatization of the uracil ring in UTP (1b) eliminated agonist properties and improved potency and pharmacokinetic properties, respectively [28].…”

Section: Introductionmentioning

confidence: 99%

“…Owing to the high demand for a selective P2Y 2 receptor antagonist, we re-synthesized AR-C118925 (3) together with two derivatives, established an optimized procedure, and assessed their antagonistic potential on the P2Y 2 receptor and related targets, including further subtypes of the P2Y receptor family, P2X receptor ion channels, adenosine receptors (P1 receptors), enzymes involved in nucleotide metabolism (ectonucleotidases), as well as a range of other potential targets that complement the selectivity testing previously published by Kemp et al [27]. Comprehensive in vitro evaluation of AR-C118925 (3) provided in the present study, including physicochemical, pharmacokinetic, and pharmacological data will provide a basis for future in vivo studies.…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Synthesis, characterization, and in vitro evaluation of the selective P2Y2 receptor antagonist AR-C118925

Rafehi

Burbiel

Attah

et al. 2016

Purinergic Signalling

View full text Add to dashboard Cite

The G q protein-coupled, ATP-and UTP-activated P2Y 2 receptor is a potential drug target for a range of different disorders, including tumor metastasis, inflammation, atherosclerosis, kidney disorders, and osteoporosis, but pharmacological studies are impeded by the limited availability of suitable antagonists. One of the most potent and selective antagonists is the thiouracil derivative AR-C118925. However, this compound was until recently not commercially available and little is known about its properties. We therefore developed an improved procedure for the synthesis of AR-C118925 and two derivatives to allow up-scaling and assessed their potency in calcium mobilization assays on the human and rat P2Y 2 receptors recombinantly expressed in 1321N1 astrocytoma cells. The compound was further evaluated for inhibition of P2Y 2 receptor-induced β-arrestin translocation. AR-C118925 behaved as a competitive antagonist with pA 2 values of 37.2 nM (calcium assay) and 51.3 nM (β-arrestin assay). Selectivity was assessed vs. related receptors including P2X, P2Y, and adenosine receptor subtypes, as well as ectonucleotidases. AR-C118925 showed at least 50-fold selectivity against the other investigated targets, except for the P2X1 and P2X3 receptors which were blocked by AR-

show abstract

Section: Pharmacological Evaluation Of Ar-c118925 (3) and Its Derivatmentioning

confidence: 99%

Section: Pharmacological Evaluation Of Ar-c118925 (3) and Its Derivatmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Synthesis, characterization, and in vitro evaluation of the selective P2Y2 receptor antagonist AR-C118925

Rafehi

Burbiel

Attah

et al. 2016

Purinergic Signalling

View full text Add to dashboard Cite

show abstract

“…These G␣ subunits share over 80% homology in their amino acid sequences (45), hence their ability to inhibit the channel might reflect a promiscuous capacity to activate the same downstream signaling pathways, one of which inhibits ENaC. Gq/11-coupled GPCRs are known to regulate several cellular events in epithelial cells, including intracellular Ca 2ϩ mobilization (46), mucin secretion (47,48), and anion secretion (49). Our data suggest that, Gq/11 plays a small, but significant, part in the P2Y 2 receptor-mediated signaling mechanism that inhibits activity of ENaC.…”

Section: Discussionmentioning

confidence: 99%

Regulation of the Epithelial Na+ Channel by the RH Domain of G Protein-coupled Receptor Kinase, GRK2, and Gαq/11

Lee

Song

Campbell

et al. 2011

Journal of Biological Chemistry

View full text Add to dashboard Cite

The G protein-coupled receptor kinase (GRK2) belongs to a family of protein kinases that phosphorylates agonist-activated G protein-coupled receptors, leading to G protein-receptor uncoupling and termination of G protein signaling. GRK2 also contains a regulator of G protein signaling homology (RH) domain, which selectively interacts with ␣-subunits of the Gq/11 family that are released during G protein-coupled receptor activation. We have previously reported that kinase activity of GRK2 up-regulates activity of the epithelial sodium channel (ENaC) in a Na ؉ absorptive epithelium by blocking Nedd4-2-dependent inhibition of ENaC. In the present study, we report that GRK2 also regulates ENaC by a mechanism that does not depend on its kinase activity. We show that a wild-type GRK2 (wtGRK2) and a kinase-dead GRK2 mutant ( K220R GRK2), but not a GRK2 mutant that lacks the C-terminal RH domain (⌬RH-GRK2) or a GRK2 mutant that cannot interact with G␣q/11/14 ( D110A GRK2), increase activity of ENaC. GRK2 up-regulates the basal activity of the channel as a consequence of its RH domain binding the ␣-subunits of Gq/11. We further found that expression of constitutively active G␣q/11 mutants significantly inhibits activity of ENaC. Conversely, co-expression of siRNA against G␣q/11 increases ENaC activity. The effect of G␣q on ENaC activity is not due to change in ENaC membrane expression and is independent of Nedd4-2. These findings reveal a novel mechanism by which GRK2 and Gq/11 ␣-subunits regulate the activity ENaC.The ␤-adrenergic receptor kinase 1 (GRK2) is one of the G protein-coupled receptor serine/threonine kinases (GRKs). 3All seven members of the GRK family (GRK1-7) share a highly homologous kinase domain that is flanked toward the C-terminal by a pleckstrin homology (PH) domain and toward the N-terminal by a regulator of G-protein signaling homology (RH) domain (1). Protein kinases of this family are unique in their ability to specifically phosphorylate the agonist-activated form of heptahelical G protein-coupled receptors (GPCRs) (2). Upon stimulation, GRKs facilitate binding of agonist-activated GPCRs to cytosolic cofactor proteins, arrestins, and other proteins involved in receptor desensitization (3,4). This interaction impairs coupling between the GPCRs and trimeric G proteins, targets GPCRs for clathrin-mediated endocytosis (3), and terminates G protein signal transduction (2). Activity of GRKs is, therefore, important for arbitrating an appropriate strength and duration of cellular responses, allowing the G protein-dependent cellular responses to physiological stimulation to cease rapidly after receptor activation even in the continuing presence of stimuli.Recent studies suggest that, in addition to their ability to inactivate GPCRs by phosphorylation, GRKs can phosphorylate and regulate activity of an array of non-receptor substrates (2) and also regulate GPCR signaling by a mechanism that does not require their intrinsic kinase activity (3). For instance, the RH domain of GRK2 contains a binding site th...

show abstract

P2Y₂ receptor antagonists as anti‐allodynic agents in acute and sub‐chronic trigeminal sensitization: Role of satellite glial cells

Magni

Merli

Verderio

et al. 2015

Glia

View full text Add to dashboard Cite

Trigeminal (TG) pain often lacks a satisfactory pharmacological control. A better understanding of the molecular cross-talk between TG neurons and surrounding satellite glial cells (SGCs) could help identifying innovative targets for the development of more effective analgesics. We have previously demonstrated that neuronal pro-algogenic mediators upregulate G protein-coupled nucleotide P2Y receptors (P2YRs) expressed by TG SGCs in vitro. Here, we have identified the specific P2YR subtypes involved (i.e., the ADP-sensitive P2Y1 R and the UTP-responsive P2Y2 R subtypes), and demonstrated the contribution of neuron-derived prostaglandins to their upregulation. Next, we have translated these data to an in vivo model of TG pain (namely, rats injected with Complete Freund's adjuvant in the temporomandibular joint), by demonstrating activation of SGCs and upregulation of P2Y1 R and P2Y2 R in the ipsi-lateral TG. To unequivocally link P2YRs to the development of facial allodynia, we treated animals with various purinergic antagonists. The selective P2Y2 R antagonist AR-C118925 completely inhibited SGCs activation, exerted a potent anti-allodynic effect that lasted over time, and was still effective when administration was started 6-days post induction of allodynia, i.e. under subchronic pain conditions. Conversely, the selective P2Y1 R antagonist MRS2179 was completely ineffective. Moreover, similarly to the anti-inflammatory drug acetylsalicylic acid and the known anti-migraine agent sumatriptan, the P2X/P2Y nonselective antagonist PPADS was only partially effective, and completely lost its activity under sub-chronic conditions. Taken together, our results highlight glial P2Y2 Rs as potential "druggable" targets for the successful management of TG-related pain.

show abstract

Nucleotide-Mediated Mucin Secretion from Differentiated Human Bronchial Epithelial Cells

Cited by 82 publications

References 34 publications

Synthesis, characterization, and in vitro evaluation of the selective P2Y2 receptor antagonist AR-C118925

Synthesis, characterization, and in vitro evaluation of the selective P2Y2 receptor antagonist AR-C118925

Regulation of the Epithelial Na+ Channel by the RH Domain of G Protein-coupled Receptor Kinase, GRK2, and Gαq/11

P2Y₂ receptor antagonists as anti‐allodynic agents in acute and sub‐chronic trigeminal sensitization: Role of satellite glial cells

Contact Info

Product

Resources

About

Nucleotide-Mediated Mucin Secretion from Differentiated Human Bronchial Epithelial Cells

Cited by 82 publications

References 34 publications

Synthesis, characterization, and in vitro evaluation of the selective P2Y2 receptor antagonist AR-C118925

Synthesis, characterization, and in vitro evaluation of the selective P2Y2 receptor antagonist AR-C118925

Regulation of the Epithelial Na+ Channel by the RH Domain of G Protein-coupled Receptor Kinase, GRK2, and Gαq/11

P2Y2 receptor antagonists as anti‐allodynic agents in acute and sub‐chronic trigeminal sensitization: Role of satellite glial cells

Contact Info

Product

Resources

About

P2Y₂ receptor antagonists as anti‐allodynic agents in acute and sub‐chronic trigeminal sensitization: Role of satellite glial cells