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Pinacidil, a Katp channel opener, identified as a novel agonist for TRPA1

Abstract

The transient receptor potential Ankyrin 1 (TRPA1) cation channel is activated by various pungent and irritant compounds, and it also mediates the perception of noxious cold. Identification of different agonists for this channel is important for understanding its activation mechanism. Therefore, a screen for novel TRPA1 agonists was performed using an agonist-induced calcium influx assay. Out of 90 compounds screened, pinacidil was identified as a novel agonist for this channel. Pinacidil is a known opener of the Katp channel, for which it has an EC50 value of 1–3 μmol/L. In comparison, the EC50 value of pinacidil for TRPA1 is relatively high (260 μmol/L). Recombinant HEK-TRPA1 cells did not respond to P1075, another Katp channel opener, suggesting that the effect of pinacidil on TRPA1 was highly specific. Further studies revealed that the agonist activity of pinacidil could be blocked by the TRP channel inhibitors, ruthenium red and HC-030031. Using glutathione (GSH) and site-specific mutagenesis, we demonstrated that pinacidil could activate TRPA1 by covalent modification of the critical amino acids C619, C639 and C663 in the N-terminus of TRPA1.

References

  1. Patapoutian A, Peier A M, Story G M, et al. ThermoTRP channels and beyond: Mechanisms of temperature sensation. Nat Rev Neurosci, 2003, 4: 529–539

    Article  Google Scholar 

  2. Alexander S P, Mathie A, Peters J A. Guide to receptors and channels. 2nd ed. Br J Pharmacol, 2006, 147(Suppl 3): S1–168

    Google Scholar 

  3. Kwan K Y, Allchorne A J, Vollrath M A, et al. TRPA1 contributes to cold, mechanical, and chemical nociception but is not essential for hair-cell transduction. Neuron, 2006, 50: 277–289

    Article  Google Scholar 

  4. Vannier B, Zhu X, Brown D, et al. The membrane topology of human transient receptor potential 3 as inferred from glycosylation-scanning mutagenesis and epitope immunocytochemistry. J Biol Chem, 1998, 273: 8675–8679

    Article  Google Scholar 

  5. Montell C. Thermosensation: Hot findings make TRPNs very cool. Curr Biol, 2003, 13: R476–478

    Article  Google Scholar 

  6. Nilius B, Owsianik G, Voets T, et al. Transient receptor potential cation channels in disease. Physiol Rev, 2007, 87: 165–217

    Article  Google Scholar 

  7. Ramsey I S, Delling M, Clapham D E. An introduction to TRP channels. Annu Rev Physiol, 2006, 68: 619–647

    Article  Google Scholar 

  8. Jaquemar D, Schenker T, Trueb B. An ankyrin-like protein with transmembrane domains is specifically lost after oncogenic transformation of human fibroblasts. J Biol Chem, 1999, 274: 7325–7333

    Article  Google Scholar 

  9. Story G M, Peier A M, Reeve A J, et al. ANKTM1, a TRP-like channel expressed in nociceptive neurons, is activated by cold temperatures. Cell, 2003, 112: 819–829

    Article  Google Scholar 

  10. Nagata K, Duggan A, Kumar G, et al. Nociceptor and hair cell transducer properties of TRPA1, a channel for pain and hearing. J Neurosci, 2005, 25: 4052–4061

    Article  Google Scholar 

  11. Stokes A, Wakano C, Koblan-Huberson M, et al. TRPA1 is a substrate for de-ubiquitination by the tumor suppressor CYLD. Cell Signal, 2006, 18: 1584–1594

    Article  Google Scholar 

  12. Nozawa K, Kawabata-Shoda E, Doihara H, et al. TRPA1 regulates gastrointestinal motility through serotonin release from enterochromaffin cells. Proc Natl Acad Sci USA, 2009, 106: 3408–3413

    Article  Google Scholar 

  13. Bandell M, Story G M, Hwang S W, et al. Noxious cold ion channel TRPA1 is activated by pungent compounds and bradykinin. Neuron, 2004, 41: 849–857

    Article  Google Scholar 

  14. Baraldi P G, Preti D, Materazzi S, et al. Transient receptor potential ankyrin 1 (TRPA1) channel as emerging target for novel analgesics and anti-inflammatory agents. J Med Chem, 2010, 53: 5085–5107

    Article  Google Scholar 

  15. Bautista D M, Movahed P, Hinman A, et al. Pungent products from garlic activate the sensory ion channel TRPA1. Proc Natl Acad Sci USA, 2005, 102: 12248–12252

    Article  Google Scholar 

  16. Bautista D M, Jordt S E, Nikai T, et al. TRPA1 mediates the inflammatory actions of environmental irritants and proalgesic agents. Cell, 2006, 124: 1269–1282

    Article  Google Scholar 

  17. Terada Y, Narukawa M, Watanabe T. Specific hydroxy fatty acids in royal jelly activate TRPA1. J Agric Food Chem, 2011, 59: 2627–2635

    Article  Google Scholar 

  18. Gijsen H J, Berthelot D, Zaja M, et al. Analogues of morphanthridine and the tear gas dibenz[b,f][1,4]oxazepine (CR) as extremely potent activators of the human transient receptor potential ankyrin 1 (TRPA1) channel. J Med Chem, 2010, 53: 7011–7020

    Article  Google Scholar 

  19. Andre E, Gatti R, Trevisani M, et al. Transient receptor potential ankyrin receptor 1 is a novel target for pro-tussive agents. Br J Pharmacol, 2009, 158: 1621–1628

    Article  Google Scholar 

  20. Nilius B. From TRPs to SOCs, CCEs, and CRACs: Consensus and controversies. Cell Calcium, 2003, 33: 293–298

    Article  Google Scholar 

  21. Rankow R M. Question: What is the best method to deal with bony ankylosis of the temporomandibular joint? Ann Plast Surg, 1978, 1: 627

    Article  Google Scholar 

  22. Lin Y F, Raab-Graham K, Jan Y N, et al. NO stimulation of ATP-sensitive potassium channels: Involvement of Ras/mitogen-activated protein kinase pathway and contribution to neuroprotection. Proc Natl Acad Sci USA, 2004, 101: 7799–7804

    Article  Google Scholar 

  23. Sun X, Cao K, Yang G, et al. Selective expression of Kir6.1 protein in different vascular and non-vascular tissues. Biochem Pharmacol, 2004, 67: 147–156

    Google Scholar 

  24. Prost A, Derand R, Gros L, et al. Inhibition of ATP-sensitive K+ channels by substituted benzo[c]quinolizinium CFTR activators. Biochem Pharmacol, 2003, 66: 425–430

    Article  Google Scholar 

  25. Gojkovic-Bukarica L C, Beleslin-Cokic B B, Novakovic A N, et al. The effects of potassium channel opener P1075 on the human saphenous vein and human internal mammary artery. J Cardiovasc Pharmacol, 2011 (in press)

  26. Hinman A, Chuang H H, Bautista D M, et al. TRP channel activation by reversible covalent modification. Proc Natl Acad Sci USA, 2006, 103: 19564–19568

    Article  Google Scholar 

  27. Jordt S E, Bautista D M, Chuang H H, et al. Mustard oils and cannabinoids excite sensory nerve fibres through the TRP channel ANKTM1. Nature, 2004, 427: 260–265

    Article  Google Scholar 

  28. Eid S R, Crown E D, Moore E L, et al. HC-030031, a TRPA1 selective antagonist, attenuates inflammatory- and neuropathy-induced mechanical hypersensitivity. Mol Pain, 2008, 4: 48

    Article  Google Scholar 

  29. Taylor-Clark T E, Undem B J, Macglashan D W, et al. Prostaglandin-induced activation of nociceptive neurons via direct interaction with transient receptor potential A1 (TRPA1). Mol Pharmacol, 2008, 73: 274–281

    Article  Google Scholar 

  30. Maher M, Ao H, Banke T, et al. Activation of TRPA1 by farnesyl thiosalicylic acid. Mol Pharmacol, 2008, 73: 1225–1234

    Article  Google Scholar 

  31. Doerner J F, Gisselmann G, Hatt H, et al. Transient receptor potential channel A1 is directly gated by calcium ions. J Biol Chem, 2007, 282: 13180–13189

    Article  Google Scholar 

  32. Zurborg S, Yurgionas B, Jira J A, et al. Direct activation of the ion channel TRPA1 by Ca2+. Nat Neurosci, 2007, 10: 277–279

    Article  Google Scholar 

  33. Cavanaugh E J, Simkin D, Kim D. Activation of transient receptor potential A1 channels by mustard oil, tetrahydrocannabinol and Ca2+ reveals different functional channel states. Neuroscience, 2008, 154: 1467–1476

    Article  Google Scholar 

  34. Macpherson L J, Dubin A E, Evans M J, et al. Noxious compounds activate TRPA1 ion channels through covalent modification of cysteines. Nature, 2007, 445: 541–545

    Article  Google Scholar 

  35. Sterndorff B, Johansen P. The antihypertensive effect of pinacidil versus prazosin in mild to moderate hypertensive patients seen in general practice. Acta Med Scand, 1988, 224: 329–336

    Article  Google Scholar 

  36. Friedel H A, Brogden R N. Pinacidil. A review of its pharmacodynamic and pharmacokinetic properties, and therapeutic potential in the treatment of hypertension. Drugs, 1990, 39: 929–967

    Article  Google Scholar 

  37. Obata K, Katsura H, Mizushima T, et al. TRPA1 induced in sensory neurons contributes to cold hyperalgesia after inflammation and nerve injury. J Clin Invest, 2005, 115: 2393–2401

    Article  Google Scholar 

  38. Caterina M J, Leffler A, Malmberg A B, et al. Impaired nociception and pain sensation in mice lacking the capsaicin receptor. Science, 2000, 288: 306–313

    Article  Google Scholar 

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Correspondence to JianFeng Liu.

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Ma, L., Deng, Y., Zhang, B. et al. Pinacidil, a Katp channel opener, identified as a novel agonist for TRPA1. Chin. Sci. Bull. 57, 1810–1817 (2012). https://doi.org/10.1007/s11434-012-5035-0

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  • DOI: https://doi.org/10.1007/s11434-012-5035-0

Keywords

  • TRPA1
  • pinacidil
  • covalent modification