The ankyrin transient receptor potential channel TRPA1 is a non-selective cationic channel that is expressed by sensory neurons, where it can be activated by pungent chemicals, such as AITC (allyl isothiocyanate), cinnamon or allicin, by deep cooling (<18 °C) or highly depolarizing voltages (>+100 mV). From the cytoplasmic side, this channel can be regulated by negatively charged ligands such as phosphoinositides or inorganic polyphosphates, most likely through an interaction with as yet unidentified positively charged domain(s). In the present study, we mutated 27 basic residues along the C-terminal tail of TRPA1, trying to explore their role in AITC- and voltage-dependent gating. In the proximal part of the C-terminus, the function-affecting mutations were at Lys969, Arg975, Lys988 and Lys989. A second significant region was found in the predicted helix, centred around Lys1048 and Lys1052, in which single alanine mutations completely abolished AITC- and voltage-dependent activation. In the distal portion of the C-terminus, the charge neutralizations K1092A and R1099A reduced the AITC sensitivity, and, in the latter mutant, increased the voltage-induced steady-state responses. Taken together, our findings identify basic residues in the C-terminus that are strongly involved in TRPA1 voltage and chemical sensitivity, and some of them may represent possible interaction sites for negatively charged molecules that are generally considered to modulate TRPA1.

Department of Cellular Neurophysiology Institute of Physiology Academy of Sciences of the Czech Republic Videnska 1083 142 20 Prague 4 Czech Republic

Zobrazit více v PubMed

Story G. M., Peier A. M., Reeve A. J., Eid S. R., Mosbacher J., Hricik T. R., Earley T. J., Hergarden A. C., Andersson D. A., Hwang S. W., et al. ANKTM1, a TRP-like channel expressed in nociceptive neurons, is activated by cold temperatures. Cell. 2003;112:819–829. PubMed

Zhang X. F., Chen J., Faltynek C. R., Moreland R. B., Neelands T. R. Transient receptor potential A1 mediates an osmotically activated ion channel. Eur. J. Neurosci. 2008;27:605–611. PubMed

Sawada Y., Hosokawa H., Hori A., Matsumura K., Kobayashi S. Cold sensitivity of recombinant TRPA1 channels. Brain Res. 2007;1160:39–46. PubMed

Macpherson L. J., Dubin A. E., Evans M. J., Marr F., Schultz P. G., Cravatt B. F., Patapoutian A. Noxious compounds activate TRPA1 ion channels through covalent modification of cysteines. Nature. 2007;445:541–545. PubMed

Hinman A., Chuang H. H., Bautista D. M., Julius D. TRP channel activation by reversible covalent modification. Proc. Natl. Acad. Sci. U.S.A. 2006;103:19564–19568. PubMed PMC

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. PubMed

Garcia-Anoveros J., Nagata K. TRPA1. Handb. Exp. Pharmacol. 2007;179:347–362. PubMed

Bautista D. M., Jordt S. E., Nikai T., Tsuruda P. R., Read A. J., Poblete J., Yamoah E. N., Basbaum A. I., Julius D. TRPA1 mediates the inflammatory actions of environmental irritants and proalgesic agents. Cell. 2006;124:1269–1282. PubMed

Nagata K., Duggan A., Kumar G., Garcia-Anoveros J. Nociceptor and hair cell transducer properties of TRPA1, a channel for pain and hearing. J. Neurosci. 2005;25:4052–4061. PubMed PMC

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

Zurborg S., Yurgionas B., Jira J. A., Caspani O., Heppenstall P. A. Direct activation of the ion channel TRPA1 by Ca2+ Nat. Neurosci. 2007;10:277–279. PubMed

Wang Y. Y., Chang R. B., Waters H. N., McKemy D. D., Liman E. R. The nociceptor ion channel TRPA1 is potentiated and inactivated by permeating calcium ions. J. Biol. Chem. 2008;283:32691–32703. PubMed PMC

Bandell M., Story G. M., Hwang S. W., Viswanath V., Eid S. R., Petrus M. J., Earley T. J., Patapoutian A. Noxious cold ion channel TRPA1 is activated by pungent compounds and bradykinin. Neuron. 2004;41:849–857. PubMed

Wang S., Dai Y., Fukuoka T., Yamanaka H., Kobayashi K., Obata K., Cui X., Tominaga M., Noguchi K. Phospholipase C and protein kinase A mediate bradykinin sensitization of TRPA1: a molecular mechanism of inflammatory pain. Brain. 2008;131:1241–1251. PubMed

Nilius B., Owsianik G., Voets T. Transient receptor potential channels meet phosphoinositides. EMBO J. 2008;27:2809–2816. PubMed PMC

Corey D. P., Garcia-Anoveros J., Holt J. R., Kwan K. Y., Lin S. Y., Vollrath M. A., Amalfitano A., Cheung E. L., Derfler B. H., Duggan A., et al. TRPA1 is a candidate for the mechanosensitive transduction channel of vertebrate hair cells. Nature. 2004;432:723–730. PubMed

Hirono M., Denis C. S., Richardson G. P., Gillespie P. G. Hair cells require phosphatidylinositol 4,5-bisphosphate for mechanical transduction and adaptation. Neuron. 2004;44:309–320. PubMed

Karashima Y., Prenen J., Meseguer V., Owsianik G., Voets T., Nilius B. Modulation of the transient receptor potential channel TRPA1 by phosphatidylinositol 4,5-biphosphate manipulators. Pflugers Arch. 2008;457:77–89. PubMed

Akopian A. N., Ruparel N. B., Jeske N. A., Hargreaves K. M. Transient receptor potential TRPA1 channel desensitization in sensory neurons is agonist dependent and regulated by TRPV1-directed internalization. J. Physiol. 2007;583:175–193. PubMed PMC

Kim D., Cavanaugh E., Simkin D. Inhibition of transient receptor potential A1 by phosphatidylinositol-4,5-bisphosphate. Am. J. Physiol. Cell Physiol. 2008;295:C92–C99. PubMed PMC

Rohacs T. Phosphoinositide regulation of non-canonical transient receptor potential channels. Cell Calcium. 2009;45:554–565. PubMed PMC

Suh B. C., Hille B. PIP2 is a necessary cofactor for ion channel function: how and why? Annu. Rev. Biophys. 2008;37:175–195. PubMed PMC

Karashima Y., Talavera K., Everaerts W., Janssens A., Kwan K. Y., Vennekens R., Nilius B., Voets T. TRPA1 acts as a cold sensor in vitro and in vivo. Proc. Natl. Acad. Sci. U.S.A. 2009;106:1273–1278. PubMed PMC

Susankova K., Ettrich R., Vyklicky L., Teisinger J., Vlachova V. Contribution of the putative inner-pore region to the gating of the transient receptor potential vanilloid subtype 1 channel (TRPV1) J. Neurosci. 2007;27:7578–7585. PubMed PMC

Vlachova V., Teisinger J., Sušánková K., Lyfenko A., Ettrich R., Vyklicky L. Functional role of C-terminal cytoplasmic tail of rat vanilloid receptor 1. J. Neurosci. 2003;23:1340–1350. PubMed PMC

Dittert I., Benedikt J., Vyklicky L., Zimmermann K., Reeh P. W., Vlachova V. Improved superfusion technique for rapid cooling or heating of cultured cells under patch-clamp conditions. J. Neurosci. Methods. 2006;151:178–185. PubMed

Benedikt J., Samad A., Ettrich R., Teisinger J., Vlachova V. Essential role for the putative S6 inner pore region in the activation gating of the human TRPA1 channel. Biochim. Biophys. Acta. 2009;1793:1279–1288. PubMed

Andersson D. A., Gentry C., Moss S., Bevan S. Transient receptor potential A1 is a sensory receptor for multiple products of oxidative stress. J. Neurosci. 2008;28:2485–2494. PubMed PMC

Karashima Y., Damann N., Prenen J., Talavera K., Segal A., Voets T., Nilius B. Bimodal action of menthol on the transient receptor potential channel TRPA1. J. Neurosci. 2007;27:9874–9884. PubMed PMC

Latorre R., Zaelzer C., Brauchi S. Structure-functional intimacies of transient receptor potential channels. Q. Rev. Biophys. 2009;42:201–246. PubMed

Matta J. A., Ahern G. P. Voltage is a partial activator of rat thermosensitive TRP channels. J. Physiol. 2007;585:469–482. PubMed PMC

Jordt S. E., Julius D. Molecular basis for species-specific sensitivity to “hot” chili peppers. Cell. 2002;108:421–430. PubMed

Gavva N. R., Klionsky L., Qu Y., Shi L., Tamir R., Edenson S., Zhang T. J., Viswanadhan V. N., Toth A., Pearce L. V., et al. Molecular determinants of vanilloid sensitivity in TRPV1. J. Biol. Chem. 2004;279:20283–20295. PubMed

Brauchi S., Orta G., Mascayano C., Salazar M., Raddatz N., Urbina H., Rosenmann E., Gonzalez-Nilo F., Latorre R. Dissection of the components for PIP2 activation and thermosensation in TRP channels. Proc. Natl. Acad. Sci. U.S.A. 2007;104:10246–10251. PubMed PMC

Hu H., Grandl J., Bandell M., Petrus M., Patapoutian A. Two amino acid residues determine 2-APB sensitivity of the ion channels TRPV3 and TRPV4. Proc. Natl. Acad. Sci. U.S.A. 2009;106:1626–1631. PubMed PMC

Grandl J., Hu H., Bandell M., Bursulaya B., Schmidt M., Petrus M., Patapoutian A. Pore region of TRPV3 ion channel is specifically required for heat activation. Nat. Neurosci. 2008;11:1007–1013. PubMed PMC

Yang F., Cui Y., Wang K., Zheng J. Thermosensitive TRP channel pore turret is part of the temperature activation pathway. Proc. Natl. Acad. Sci. U.S.A. 2010;107:7083–7088. PubMed PMC

Grandl J., Kim S. E., Uzzell V., Bursulaya B., Petrus M., Bandell M., Patapoutian A. Temperature-induced opening of TRPV1 ion channel is stabilized by the pore domain. Nat. Neurosci. 2010;13:708–714. PubMed PMC

Loukin S., Su Z., Zhou X., Kung C. Forward-genetic analysis reveals multiple gating mechanisms of Trpv4. J. Biol. Chem. 2010;285:19884–19890. PubMed PMC

Myers B. R., Bohlen C. J., Julius D. A yeast genetic screen reveals a critical role for the pore helix domain in TRP channel gating. Neuron. 2008;58:362–373. PubMed PMC

Ryu S., Liu B., Yao J., Fu Q., Qin F. Uncoupling proton activation of vanilloid receptor TRPV1. J. Neurosci. 2007;27:12797–12807. PubMed PMC

Human Transient Receptor Potential Ankyrin 1 Channel: Structure, Function, and Physiology

Proximal C-Terminus Serves as a Signaling Hub for TRPA1 Channel Regulation via Its Interacting Molecules and Supramolecular Complexes

Regulation of the transient receptor potential channel TRPA1 by its N-terminal ankyrin repeat domain

C-terminal acidic cluster is involved in Ca2+-induced regulation of human transient receptor potential ankyrin 1 channel