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. 2013 Nov 18;4(12):942–950. doi: 10.1007/s13238-013-3091-0

Crystal structure of the N-terminal ankyrin repeat domain of TRPV3 reveals unique conformation of finger 3 loop critical for channel function

Di-Jing Shi 1, Sheng Ye 2, Xu Cao 1, Rongguang Zhang 2,✉, KeWei Wang 1,3,4,✉
PMCID: PMC4875401  PMID: 24248473

Abstract

In all six members of TRPV channel subfamily, there is an ankyrin repeat domain (ARD) in their intracellular Ntermini. Ankyrin (ANK) repeat, a common motif with typically 33 residues in each repeat, is primarily involved in protein-protein interactions. Despite the sequence similarity among the ARDs of TRPV channels, the structure of TRPV3-ARD, however, remains unknown. Here, we report the crystal structure of TRPV3-ARD solved at 1.95 Å resolution, which reveals six-ankyrin repeats. While overall structure of TRPV3-ARD is similar to ARDs from other members of TRPV subfamily; it, however, features a noticeable finger 3 loop that bends over and is stabilized by a network of hydrogen bonds and hydrophobic packing, instead of being flexible as seen in known TRPV-ARD structures. Electrophysiological recordings demonstrated that mutating key residues R225, R226, Q255, and F249 of finger 3 loop altered the channel activities and pharmacology. Taken all together, our findings show that TRPV3-ARD with characteristic finger 3 loop likely plays an important role in channel function and pharmacology.

Keywords: TRPV3, ARD, keratinocyte, 2-APB, skin

Footnotes

These authors contributed equally to the work.

Contributor Information

Rongguang Zhang, Email: rzhang@ibp.ac.cn.

KeWei Wang, Email: wangkw@hsc.pku.edu.cn.

References

  1. Adams PD, Afonine PV, Bunkoczi G, Chen VB, Davis IW, Echols N, Headd JJ, Hung LW, Kapral GJ, Grosse-Kunstleve RW, et al. PHENIX: a comprehensive Python-based system for macromolecular structure solution. Acta Crystallogr D Biol Crystallogr. 2010;66:213–221. doi: 10.1107/S0907444909052925. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Arniges M, Fernandez-Fernandez JM, Albrecht N, Schaefer M, Valverde MA. Human TRPV4 channel splice variants revealed a key role of ankyrin domains in multimerization and trafficking. J Biol Chem. 2006;281:1580–1586. doi: 10.1074/jbc.M511456200. [DOI] [PubMed] [Google Scholar]
  3. Auer-Grumbach M, Olschewski A, Papic L, Kremer H, McEntagart ME, Uhrig S, Fischer C, Frohlich E, Balint Z. Alterations in the ankyrin domain of TRPV4 cause congenital distal SMA, scapuloperoneal SMA and HMSN2C. Nat Genet. 2010;42:160–164. doi: 10.1038/ng.508. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bang S, Yoo S, Yang TJ, Cho H, Hwang SW. Farnesyl pyrophosphate is a novel pain-producing molecule via specific activation of TRPV3. J Biol Chem. 2010;285:19362–19371. doi: 10.1074/jbc.M109.087742. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Caterina MJ. Transient receptor potential ion channels as participants in thermosensation and thermoregulation. Am J Physiol Regul Integr Comp Physiol. 2007;292:R64–76. doi: 10.1152/ajpregu.00446.2006. [DOI] [PubMed] [Google Scholar]
  6. Chang Q, Gyftogianni E, van de Graaf SF, Hoefs S, Weidema FA, Bindels RJ, Hoenderop JG. Molecular determinants in TRPV5 channel assembly. J Biol Chem. 2004;279:54304–54311. doi: 10.1074/jbc.M406222200. [DOI] [PubMed] [Google Scholar]
  7. Cheng X, Jin J, Hu L, Shen D, Dong XP, Samie MA, Knoff J, Eisinger B, Liu ML, Huang SM, et al. TRP channel regulates EGFR Signaling in Hair Morphogenesis and Skin Barrier Formation. Cell. 2010;141:331–343. doi: 10.1016/j.cell.2010.03.013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chung MK, Lee H, Mizuno A, Suzuki M, Caterina MJ. 2-aminoethoxydiphenyl borate activates and sensitizes the heat-gated ion channel TRPV3. J Neurosci. 2004;24:5177–5182. doi: 10.1523/JNEUROSCI.0934-04.2004. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chung MK, Lee H, Mizuno A, Suzuki M, Caterina MJ. TRPV3 and TRPV4 mediate warmth-evoked currents in primary mouse keratinocytes. J Biol Chem. 2004;279:21569–21575. doi: 10.1074/jbc.M401872200. [DOI] [PubMed] [Google Scholar]
  10. Clapham DE. TRP channels as cellular sensors. Nature. 2003;426:517–524. doi: 10.1038/nature02196. [DOI] [PubMed] [Google Scholar]
  11. Dhaka A, Uzzell V, Dubin AE, Mathur J, Petrus M, Bandell M, Patapoutian A. TRPV1 is activated by both acidic and basic pH. J Neurosci. 2009;29:153–158. doi: 10.1523/JNEUROSCI.4901-08.2009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Emsley P, Cowtan K. Coot: model-building tools for molecular graphics. Acta Crystallogr D. 2004;60:2126–2132. doi: 10.1107/S0907444904019158. [DOI] [PubMed] [Google Scholar]
  13. Erler I, Hirnet D, Wissenbach U, Flockerzi V, Niemeyer BA. Ca2+-selective transient receptor potential V channel architecture and function require a specific ankyrin repeat. J Biol Chem. 2004;279:34456–34463. doi: 10.1074/jbc.M404778200. [DOI] [PubMed] [Google Scholar]
  14. Guler AD, Lee H, Iida T, Shimizu I, Tominaga M, Caterina M. Heat-evoked activation of the ion channel, TRPV4. J Neurosci. 2002;22:6408–6414. doi: 10.1523/JNEUROSCI.22-15-06408.2002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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. doi: 10.1073/pnas.0812209106. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hu HZ, Gu Q, Wang C, Colton CK, Tang J, Kinoshita-Kawada M, Lee LY, Wood JD, Zhu MX. 2-aminoethoxydiphenyl borate is a common activator of TRPV1, TRPV2, and TRPV3. J Biol Chem. 2004;279:35741–35748. doi: 10.1074/jbc.M404164200. [DOI] [PubMed] [Google Scholar]
  17. Inada H, Procko E, Sotomayor M, Gaudet R. Structural and biochemical consequences of disease-causing mutations in the ankyrin repeat domain of the human TRPV4 channel. Biochemistry. 2012;51:6195–6206. doi: 10.1021/bi300279b. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Jin X, Touhey J, Gaudet R. Structure of the N-terminal ankyrin repeat domain of the TRPV2 ion channel. J Biol Chem. 2006;281:25006–25010. doi: 10.1074/jbc.C600153200. [DOI] [PubMed] [Google Scholar]
  19. Kim S, Kang C, Shin CY, Hwang SW, Yang YD, Shim WS, Park MY, Kim E, Kim M, Kim BM, et al. TRPV1 recapitulates native capsaicin receptor in sensory neurons in association with Fas-associated factor 1. J Neurosci. 2006;26:2403–2412. doi: 10.1523/JNEUROSCI.4691-05.2006. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Landoure G, Zdebik AA, Martinez TL, Burnett BG, Stanescu HC, Inada H, Shi Y, Taye AA, Kong L, Munns CH, et al. Mutations in TRPV4 cause Charcot-Marie-Tooth disease type 2C. Nat Genet. 2010;42:170–174. doi: 10.1038/ng.512. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lin Z, Chen Q, Lee M, Cao X, Zhang J, Ma D, Chen L, Hu X, Wang H, Wang X, et al. Exome sequencing reveals mutations in TRPV3 as a cause of Olmsted syndrome. Am J Hum Genet. 2012;90:558–564. doi: 10.1016/j.ajhg.2012.02.006. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lishko PV, Procko E, Jin X, Phelps CB, Gaudet R. The ankyrin repeats of TRPV1 bind multiple ligands and modulate channel sensitivity. Neuron. 2007;54:905–918. doi: 10.1016/j.neuron.2007.05.027. [DOI] [PubMed] [Google Scholar]
  23. McCleverty CJ, Koesema E, Patapoutian A, Lesley SA, Kreusch A. Crystal structure of the human TRPV2 channel ankyrin repeat domain. Protein Sci. 2006;15:2201–2206. doi: 10.1110/ps.062357206. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Moiseenkova-Bell VY, Stanciu LA, Serysheva II, Tobe BJ, Wensel TG. Structure of TRPV1 channel revealed by electron cryomicroscopy. Proc Natl Acad Sci U S A. 2008;105:7451–7455. doi: 10.1073/pnas.0711835105. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Montell C, Birnbaumer L, Flockerzi V. The TRP channels, a remarkably functional family. Cell. 2002;108:595–598. doi: 10.1016/S0092-8674(02)00670-0. [DOI] [PubMed] [Google Scholar]
  26. Moqrich A, Hwang SW, Earley TJ, Petrus MJ, Murray AN, Spencer KSR, Andahazy M, Story GM, Patapoutian A. Impaired thermosensation in mice lacking TRPV3, a heat and camphor sensor in the skin. Science. 2005;307:1468–1472. doi: 10.1126/science.1108609. [DOI] [PubMed] [Google Scholar]
  27. Otwinowski Z, Minor W. Processing of X-ray diffraction data collected in oscillation mode. Method Enzymol. 1997;276:307–326. doi: 10.1016/S0076-6879(97)76066-X. [DOI] [PubMed] [Google Scholar]
  28. Phelps CB, Huang RJ, Lishko PV, Wang RR, Gaudet R. Structural analyses of the ankyrin repeat domain of TRPV6 and related TRPV ion channels. Biochemistry. 2008;47:2476–2484. doi: 10.1021/bi702109w. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Phelps CB, Wang RR, Choo SS, Gaudet R. Differential regulation of TRPV1, TRPV3, and TRPV4 sensitivity through a conserved binding site on the ankyrin repeat domain. J Biol Chem. 2010;285:731–740. doi: 10.1074/jbc.M109.052548. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Shigematsu H, Sokabe T, Danev R, Tominaga M, Nagayama K. A 3.5-nm structure of rat TRPV4 cation channel revealed by Zernike phase-contrast cryoelectron microscopy. J Biol Chem. 2010;285:11210–11218. doi: 10.1074/jbc.M109.090712. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Stokes AJ, Wakano C, Del Carmen KA, Koblan-Huberson M, Turner H. Formation of a physiological complex between TRPV2 and RGA protein promotes cell surface expression of TRPV2. J Cell Biochem. 2005;94:669–683. doi: 10.1002/jcb.20331. [DOI] [PubMed] [Google Scholar]
  32. Venkatachalam K, Montell C. TRP channels. Annu Rev Biochem. 2007;76:387–417. doi: 10.1146/annurev.biochem.75.103004.142819. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Voets T, Droogmans G, Wissenbach U, Janssens A, Flockerzi V, Nilius B. The principle of temperature-dependent gating in cold- and heat-sensitive TRP channels. Nature. 2004;430:748–754. doi: 10.1038/nature02732. [DOI] [PubMed] [Google Scholar]
  34. Xiao R, Tang J, Wang C, Colton CK, Tian J, Zhu MX. Calcium plays a central role in the sensitization of TRPV3 channel to repetitive stimulations. J Biol Chem. 2008;283:6162–6174. doi: 10.1074/jbc.M706535200. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Xiao R, Tian J, Tang J, Zhu MX. The TRPV3 mutation associated with the hairless phenotype in rodents is constitutively active. Cell Calcium. 2008;43:334–343. doi: 10.1016/j.ceca.2007.06.004. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Xu H, Delling M, Jun JC, Clapham DE. Oregano, thyme and clove-derived flavors and skin sensitizers activate specific TRP channels. Nat Neurosci. 2006;9:628–635. doi: 10.1038/nn1692. [DOI] [PubMed] [Google Scholar]
  37. Yoshida T, Inoue R, Morii T, Takahashi N, Yamamoto S, Hara Y, Tominaga M, Shimizu S, Sato Y, Mori Y. Nitric oxide activates TRP channels by cysteine S-nitrosylation. Nat Chem Biol. 2006;2:596–607. doi: 10.1038/nchembio821. [DOI] [PubMed] [Google Scholar]
  38. Yoshioka T, Imura K, Asakawa M, Suzuki M, Oshima I, Hirasawa T, Sakata T, Horikawa T, Arimura A. Impact of the Gly573Ser substitution in TRPV3 on the development of allergic and pruritic dermatitis in mice. J Invest Dermatol. 2009;129:714–722. doi: 10.1038/jid.2008.245. [DOI] [PubMed] [Google Scholar]

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