Tags

Type your tag names separated by a space and hit enter

Protein kinase D and Gβγ mediate sustained nociceptive signaling by biased agonists of protease-activated receptor-2.
J Biol Chem. 2019 07 05; 294(27):10649-10662.JB

Abstract

Proteases sustain hyperexcitability and pain by cleaving protease-activated receptor-2 (PAR2) on nociceptors through distinct mechanisms. Whereas trypsin induces PAR2 coupling to Gαq, Gαs, and β-arrestins, cathepsin-S (CS) and neutrophil elastase (NE) cleave PAR2 at distinct sites and activate it by biased mechanisms that induce coupling to Gαs, but not to Gαq or β-arrestins. Because proteases activate PAR2 by irreversible cleavage, and activated PAR2 is degraded in lysosomes, sustained extracellular protease-mediated signaling requires mobilization of intact PAR2 from the Golgi apparatus or de novo synthesis of new receptors by incompletely understood mechanisms. We found here that trypsin, CS, and NE stimulate PAR2-dependent activation of protein kinase D (PKD) in the Golgi of HEK293 cells, in which PKD regulates protein trafficking. The proteases stimulated translocation of the PKD activator Gβγ to the Golgi, coinciding with PAR2 mobilization from the Golgi. Proteases also induced translocation of a photoconverted PAR2-Kaede fusion protein from the Golgi to the plasma membrane of KNRK cells. After incubation of HEK293 cells and dorsal root ganglia neurons with CS, NE, or trypsin, PAR2 responsiveness initially declined, consistent with PAR2 cleavage and desensitization, and then gradually recovered. Inhibitors of PKD, Gβγ, and protein translation inhibited recovery of PAR2 responsiveness. PKD and Gβγ inhibitors also attenuated protease-evoked mechanical allodynia in mice. We conclude that proteases that activate PAR2 by canonical and biased mechanisms stimulate PKD in the Golgi; PAR2 mobilization and de novo synthesis repopulate the cell surface with intact receptors and sustain nociceptive signaling by extracellular proteases.

Authors+Show Affiliations

From the Monash Institute of Pharmaceutical Sciences and Australian Research Council Centre of Excellence in Convergent Bio-Nano Science and Technology, Monash University, Parkville, Victoria 3052, Australia.From the Monash Institute of Pharmaceutical Sciences and Australian Research Council Centre of Excellence in Convergent Bio-Nano Science and Technology, Monash University, Parkville, Victoria 3052, Australia.the Departments of Surgery and Pharmacology, Columbia University Vagelos College of Physicians and Surgeons, Columbia University, New York, New York 10032.the Gastrointestinal Diseases Research Unit, Division of Gastroenterology, Queen's University, Kingston, Ontario K7L 3N6, Canada, and.the Departments of Surgery and Pharmacology, Columbia University Vagelos College of Physicians and Surgeons, Columbia University, New York, New York 10032.From the Monash Institute of Pharmaceutical Sciences and Australian Research Council Centre of Excellence in Convergent Bio-Nano Science and Technology, Monash University, Parkville, Victoria 3052, Australia.the Gastrointestinal Diseases Research Unit, Division of Gastroenterology, Queen's University, Kingston, Ontario K7L 3N6, Canada, and.the Gastrointestinal Diseases Research Unit, Division of Gastroenterology, Queen's University, Kingston, Ontario K7L 3N6, Canada, and.From the Monash Institute of Pharmaceutical Sciences and Australian Research Council Centre of Excellence in Convergent Bio-Nano Science and Technology, Monash University, Parkville, Victoria 3052, Australia.the Gastrointestinal Diseases Research Unit, Division of Gastroenterology, Queen's University, Kingston, Ontario K7L 3N6, Canada, and.the Bluestone Center for Clinical Research, New York University College of Dentistry, New York, New York 10010.the Departments of Surgery and Pharmacology, Columbia University Vagelos College of Physicians and Surgeons, Columbia University, New York, New York 10032, nb2733@cumc.columbia.edu.

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

31142616

Citation

Zhao, Peishen, et al. "Protein Kinase D and Gβγ Mediate Sustained Nociceptive Signaling By Biased Agonists of Protease-activated Receptor-2." The Journal of Biological Chemistry, vol. 294, no. 27, 2019, pp. 10649-10662.
Zhao P, Pattison LA, Jensen DD, et al. Protein kinase D and Gβγ mediate sustained nociceptive signaling by biased agonists of protease-activated receptor-2. J Biol Chem. 2019;294(27):10649-10662.
Zhao, P., Pattison, L. A., Jensen, D. D., Jimenez-Vargas, N. N., Latorre, R., Lieu, T., Jaramillo, J. O., Lopez-Lopez, C., Poole, D. P., Vanner, S. J., Schmidt, B. L., & Bunnett, N. W. (2019). Protein kinase D and Gβγ mediate sustained nociceptive signaling by biased agonists of protease-activated receptor-2. The Journal of Biological Chemistry, 294(27), 10649-10662. https://doi.org/10.1074/jbc.RA118.006935
Zhao P, et al. Protein Kinase D and Gβγ Mediate Sustained Nociceptive Signaling By Biased Agonists of Protease-activated Receptor-2. J Biol Chem. 2019 07 5;294(27):10649-10662. PubMed PMID: 31142616.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Protein kinase D and Gβγ mediate sustained nociceptive signaling by biased agonists of protease-activated receptor-2. AU - Zhao,Peishen, AU - Pattison,Luke A, AU - Jensen,Dane D, AU - Jimenez-Vargas,Nestor N, AU - Latorre,Rocco, AU - Lieu,TinaMarie, AU - Jaramillo,Josue O, AU - Lopez-Lopez,Cintya, AU - Poole,Daniel P, AU - Vanner,Stephen J, AU - Schmidt,Brian L, AU - Bunnett,Nigel W, Y1 - 2019/05/29/ PY - 2018/11/30/received PY - 2019/05/23/revised PY - 2019/5/31/pubmed PY - 2020/3/11/medline PY - 2019/5/31/entrez KW - F2R-like trypsin receptor KW - G protein–coupled receptor (GPCR) KW - Golgi KW - Gβγ KW - nociception KW - pain KW - protease KW - protein kinase D (PKD) KW - proteinase-activated receptor-2 KW - signal transduction KW - trafficking SP - 10649 EP - 10662 JF - The Journal of biological chemistry JO - J Biol Chem VL - 294 IS - 27 N2 - Proteases sustain hyperexcitability and pain by cleaving protease-activated receptor-2 (PAR2) on nociceptors through distinct mechanisms. Whereas trypsin induces PAR2 coupling to Gαq, Gαs, and β-arrestins, cathepsin-S (CS) and neutrophil elastase (NE) cleave PAR2 at distinct sites and activate it by biased mechanisms that induce coupling to Gαs, but not to Gαq or β-arrestins. Because proteases activate PAR2 by irreversible cleavage, and activated PAR2 is degraded in lysosomes, sustained extracellular protease-mediated signaling requires mobilization of intact PAR2 from the Golgi apparatus or de novo synthesis of new receptors by incompletely understood mechanisms. We found here that trypsin, CS, and NE stimulate PAR2-dependent activation of protein kinase D (PKD) in the Golgi of HEK293 cells, in which PKD regulates protein trafficking. The proteases stimulated translocation of the PKD activator Gβγ to the Golgi, coinciding with PAR2 mobilization from the Golgi. Proteases also induced translocation of a photoconverted PAR2-Kaede fusion protein from the Golgi to the plasma membrane of KNRK cells. After incubation of HEK293 cells and dorsal root ganglia neurons with CS, NE, or trypsin, PAR2 responsiveness initially declined, consistent with PAR2 cleavage and desensitization, and then gradually recovered. Inhibitors of PKD, Gβγ, and protein translation inhibited recovery of PAR2 responsiveness. PKD and Gβγ inhibitors also attenuated protease-evoked mechanical allodynia in mice. We conclude that proteases that activate PAR2 by canonical and biased mechanisms stimulate PKD in the Golgi; PAR2 mobilization and de novo synthesis repopulate the cell surface with intact receptors and sustain nociceptive signaling by extracellular proteases. SN - 1083-351X UR - https://www.unboundmedicine.com/medline/citation/31142616/Protein_kinase_D_and_Gβγ_mediate_sustained_nociceptive_signaling_by_biased_agonists_of_protease_activated_receptor_2_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0021-9258(20)31854-8 DB - PRIME DP - Unbound Medicine ER -