Endothelin-converting Enzyme 1 and β-Arrestins Exert Spatiotemporal Control of Substance P-induced Inflammatory Signals

Jensen, Dane D.; Halls, Michelle L.; Murphy, Jane E.; Canals, Meritxell; Cattaruzza, Fiore; Poole, Daniel P.; Lieu, Tina Marie; Koon, Hon–Wai; Pothoulakis, Charalabos; Bunnett, Nigel W.

doi:10.1074/jbc.m114.578179

Cited by 22 publications

(31 citation statements)

References 32 publications

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“…2G and fig. S3E), which also depends on βARRs and PKC but not on epidermal growth factor receptor transactivation (9, 22, 23). UBO-QIC, the phospholipase C (PLC) inhibitor U73122, and the Ca 2+ chelator EGTA prevented activation of cytosolic PKC (Fig.…”

Section: Resultsmentioning

confidence: 98%

“…We have previously shown that βARRs mediate NK 1 R endosomal signaling and nuclear ERK activation (9, 22, 23). To examine the contribution of G proteins to endosomal NK 1 R signaling, we used BRET to study SP-induced trafficking of Gα q subunits to early endosomes containing RAB5A.…”

Section: Resultsmentioning

confidence: 99%

“…8A and movie S14). By delivering activated NK 1 R to endosomes and serving as a scaffold for signaling complexes, βARRs facilitate these signals (9, 22, 23). Our findings add to the growing number of GPCRs, including β 2 -adrenergic and thyroid-stimulating hormone receptors (11, 12), known to signal from endosomes by G protein–dependent processes, and provide in vivo evidence that this endosomal mechanism is physiologically relevant.…”

Section: Discussionmentioning

confidence: 99%

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Neurokinin 1 receptor signaling in endosomes mediates sustained nociception and is a viable therapeutic target for prolonged pain relief

et al. 2017

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Typically considered to be cell surface sensors of extracellular signals, heterotrimeric GTP-binding protein (G protein)–coupled receptors (GPCRs) control many pathophysiological processes and are the target of 30% of therapeutic drugs. Activated receptors redistribute to endosomes, but researchers have yet to explore whether endosomal receptors generate signals that control complex processes in vivo and are viable therapeutic targets. We report that the substance P (SP) neurokinin 1 receptor (NK1R) signals from endosomes to induce sustained excitation of spinal neurons and pain transmission and that specific antagonism of the NK1R in endosomes with membrane-anchored drug conjugates provides more effective and sustained pain relief than conventional plasma membrane–targeted antagonists. Pharmacological and genetic disruption of clathrin, dynamin, and β-arrestin blocked SP-induced NK1R endocytosis and prevented SP-stimulated activation of cytosolic protein kinase C and nuclear extracellular signal–regulated kinase, as well as transcription. Endocytosis inhibitors prevented sustained SP-induced excitation of neurons in spinal cord slices in vitro and attenuated nociception in vivo. When conjugated to cholestanol to promote endosomal targeting, NK1R antagonists selectively inhibited endosomal signaling and sustained neuronal excitation. Cholestanol conjugation amplified and prolonged the antinociceptive actions of NK1R antagonists. These results reveal a critical role for endosomal signaling of the NK1R in the complex pathophysiology of pain and demonstrate the use of endosomally targeted GPCR antagonists.

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Section: Resultsmentioning

confidence: 98%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Neurokinin 1 receptor signaling in endosomes mediates sustained nociception and is a viable therapeutic target for prolonged pain relief

et al. 2017

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“…Measurement of [Ca 2ϩ ] i -HEK293, KNRK-PAR 2 -Kaede, or NCM460 cells were loaded with Fura-2/AM (1 M, Invitrogen) in assay buffer (150 mM NaCl, 2.6 mM KCl, 0.1 mM CaCl 2 , 1.18 mM MgCl 2 , 10 mM D-glucose, 10 mM HEPES, pH 7.4) containing 4 mM probenecid and 0.5% BSA for 1 h at 37°C (12). Fluorescence was measured at 340 and 380 nm excitation and 530 nm emission using a FlexStation III microplate reader (Molecular Devices, Sunnyvale, CA).…”

Section: Methodsmentioning

confidence: 99%

“…For some receptors, for example the substance P neurokinin 1 receptor, the calcitonin gene-related peptide receptor, and the -opioid receptor, recycling of internalized receptors is the main pathway for replenishing the plasma membrane. For these receptors, ligand dissociation and degradation in acidified early endosomes, and receptor dissociation from ␤-arrestins, are key events controlling recycling (11)(12)(13)(14)(15). Other receptors traffic to degradation pathways, and here the synthesis of new receptors or the mobilization of intracellular pools of fresh receptors is required for replenishment of the plasma membrane.…”

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confidence: 99%

Protein Kinase D and Gβγ Subunits Mediate Agonist-evoked Translocation of Protease-activated Receptor-2 from the Golgi Apparatus to the Plasma Membrane

Jensen

Zhao

Jiménez-Vargas

et al. 2016

Journal of Biological Chemistry

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Agonist-evoked endocytosis of G protein-coupled receptors has been extensively studied. The mechanisms by which agonists stimulate mobilization and plasma membrane translocation of G protein-coupled receptors from intracellular stores are unexplored. Protease-activated receptor-2 (PAR 2 ) traffics to lysosomes, and sustained protease signaling requires mobilization and plasma membrane trafficking of PAR 2 from Golgi stores. We evaluated the contribution of protein kinase D (PKD) and G␤␥ to this process. In HEK293 and KNRK cells, the PAR 2 agonists trypsin and 2-furoyl-LIGRLO-NH 2 activated PKD in the Golgi apparatus, where PKD regulates protein trafficking. PAR 2 activation induced translocation of G␤␥, a PKD activator, to the Golgi apparatus, determined by bioluminescence resonance energy transfer between G␥-Venus and giantin-Rluc8. Inhibitors of PKD (CRT0066101) and G␤␥ (gallein) prevented PAR 2 -stimulated activation of PKD. CRT0066101, PKD1 siRNA, and gallein all inhibited recovery of PAR 2 -evoked Ca 2؉ signaling. PAR 2 with a photoconvertible Kaede tag was expressed in KNRK cells to examine receptor translocation from the Golgi apparatus to the plasma membrane. Irradiation of the Golgi region (405 nm) induced green-red photo-conversion of PAR 2 -Kaede. Trypsin depleted PAR 2 -Kaede from the Golgi apparatus and repleted PAR 2 -Kaede at the plasma membrane. CRT0066101 inhibited PAR 2 -Kaede translocation to the plasma membrane. CRT0066101 also inhibited sustained protease signaling to colonocytes and nociceptive neurons that naturally express PAR 2 and mediate protease-evoked inflammation and nociception. Our results reveal a major role for PKD and G␤␥ in agonist-evoked mobilization of intracellular PAR 2 stores that is required for sustained signaling by extracellular proteases.

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miR‐20b, miR‐296, and Let‐7f Expression in Human Adipose Tissue is Related to Obesity and Type 2 Diabetes

Gentile

Lhamyani

Coín‐Aragüez

et al. 2018

Obesity

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Objective: This study aimed to analyze the potential association of different microRNA (miRNA) molecules with both type 2 diabetes (T2D) and obesity and determine their target genes. Methods: Quantitative PCR was used to analyze the miR-20b, miR-296, and Let-7f levels in human visceral and subcutaneous adipose tissues (ATs) in relation to obesity and T2D, miRTarBase 4.0 was used for validation of target genes, and the Protein Analysis Through Evolutionary Relationships (PANTHER) Classification System and the Database for Annotation, Visualization and Integrated Discovery (DAVID) were used to annotate the biological processes of the predicted targets. Results: In AT, miR-20b, miR-296, and Let-7f levels were significantly different between normoglycemic subjects and those with T2D. In visceral adipose tissue, miRNA levels were higher in normoglycemic/obesity samples than in T2D/obesity samples. miR-20b, miR-296, and Let-7f target genes that showed significant differences in both ATs in relation to obesity and T2D were CDKN1A, CX3CL1, HIF1A, PPP2R1B, STAT3, and VEGFA. These genes are known to be principally involved in the vascular endothelial growth factor (VEGF) and WNT pathways. Conclusions: This study provides experimental evidence of the possible correlation between AT miR-20b, miR-296, and Let-7f with obesity and T2D, which might involve vascular endothelial growth factor and WNTdependent pathways that are regulated by six different genes, suggesting a novel signaling pathway that could be important for understanding the mechanisms underlying the AT dysfunction associated with obesity and T2D.

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Endothelin-converting Enzyme 1 and β-Arrestins Exert Spatiotemporal Control of Substance P-induced Inflammatory Signals

Cited by 22 publications

References 32 publications

Neurokinin 1 receptor signaling in endosomes mediates sustained nociception and is a viable therapeutic target for prolonged pain relief

Neurokinin 1 receptor signaling in endosomes mediates sustained nociception and is a viable therapeutic target for prolonged pain relief

Protein Kinase D and Gβγ Subunits Mediate Agonist-evoked Translocation of Protease-activated Receptor-2 from the Golgi Apparatus to the Plasma Membrane

miR‐20b, miR‐296, and Let‐7f Expression in Human Adipose Tissue is Related to Obesity and Type 2 Diabetes

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