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Interaction of sea anemone Heteractis crispa Kunitz type polypeptides with pain vanilloid receptor TRPV1: In silico investigation

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Abstract

Using methods of molecular biology we defined the structures of the 31 sea anemone Heteractis crispa genes encoding polypeptides which are structurally homologous to the Kunitz protease inhibitor family. The identified sequences have single-point amino acid substitutions, a high degree of homology with sequences of known Kunitz family members from H. crispa, and represent a combinatorial library of polypeptides. We generated their three-dimensional structures by methods of homology modeling. Analysis of their molecular electrostatic potential allowed the division of the polypeptides into three clusters. One of them includes polypeptides APHC1, APHC2, and APHC3 which have been shown to possess, in addition to their trypsin inhibitory activity, a unique property of inhibiting the pain vanilloid receptor TRPV1 in vitro and providing the analgesic effects in vivo. The spatial structure of the polypeptide complexes with TRPV1, the nature of the interactions, as well as functionally important structural elements involved in the complex formation, were established by molecular docking technique. The designed models allowed us to propose a hypothesis contributing to the understanding of how APHC1-APHC3 affect the pain signals transduction by TRPV1: apparently, relaxation time of the receptor increases due to binding of its two chains with a polypeptide molecule which disrupts functioning of TRPV1 and leads to partial inhibition of the signal transduction in electrophysiological experiments.

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Abbreviations

APHC:

analgesic polypeptide from sea anemone Heteractis crispa

BPTI:

bovine pancreatic trypsin inhibitor

HCGS:

Gly-Ser polypeptides from sea anemone H. crispa structurally homologous to Kunitz-type protease inhibitors

MEP:

molecular electrostatic potential

SHPI-1:

Kunitz-type protease inhibitor from Caribbean sea anemone Stoichactis helianthus

TRP:

family of transient receptor potential channels

TRPV1:

pain vanilloid receptor

References

  1. Huber, R., Kukla, D., Bode, W., Schwager, P., Bartels, K., Deisenhofer, J., and Steigemann, W., J. Mol. Biol., 1974, vol. 89, pp. 73–101.

    Article  PubMed  CAS  Google Scholar 

  2. Kunitz, M. and Northrop, J., J. Gen. Physiol., 1936, vol. 19, pp. 991–1007.

    Article  PubMed  CAS  Google Scholar 

  3. Delfin, J., Martinez, I., Antuch, W., Morera, V., Gonzalez, Y., Rodriguez, R., Marquez, M., Saroyan, A., Larionova, N., Diaz, J., Padron, G., and Chavez, M., Toxicon, 1996, vol. 34, pp. 1367–1376.

    Article  PubMed  CAS  Google Scholar 

  4. Diaz, J., Morera, V., Delfin, J., Huerta, V., Lima, G., Rodriguex de la Vega, M., Garcia, B., Padron, G., Assfalg-Machleidt, I., Machleidt, W., and Chavez, M., Toxicon, 1998, vol. 36, pp. 1275–1276.

    Google Scholar 

  5. Skarzynski, T., J. Mol. Biol., 1992, vol. 224, pp. 671–683.

    Article  PubMed  CAS  Google Scholar 

  6. Foray, M.F., Lancelin, J.M., Hollecker, M., and Marion, D., Eur. J. Biochem., 1993, vol. 211, pp. 813–820.

    Article  PubMed  CAS  Google Scholar 

  7. Kunitz, M. and Northrop, J., J. Gen. Physiol., 1936, vol. 19, pp. 991–1007.

    Article  PubMed  CAS  Google Scholar 

  8. Cechova, D., in Methods in Enzymology, Lorand, L., Ed., New York: Acad. Press, 1976, vol. 45, pp. 806–813.

    Google Scholar 

  9. Ritonja, A., Meloun, B., and Gubensek, F., Biochim. Biophys. Acta, 1983, vol. 748, pp. 429–435.

    Article  PubMed  CAS  Google Scholar 

  10. Chen, C., N, Hsu C.-H., Su N.-Y., Lin Y.-C., Chiou S.-H., Wu S.-H., J. Biol. Chem., 2001, vol. 276, pp. 45079–45087.

    Article  PubMed  CAS  Google Scholar 

  11. Fritz, H., Brey, B., and Beress, L., Hoppe-Seyler’s Z. Physiol. Chem., 1972, vol. 353, pp. 19–30.

    Article  PubMed  CAS  Google Scholar 

  12. Wunderer, G., Beress, L., Machleidt, W., and Fritz, H., in Methods in Enzymology, Lorand, L., Ed., New York: Acad. Press, 1976, vol. 45, pp. 881–888.

    Google Scholar 

  13. Shiomi, K., Ishikawa, M., Yamanaka, H., and Kikuchi, T., Nippon Suisan Gakkaishi, 1989, vol. 55, pp. 1235–1241.

    Article  Google Scholar 

  14. Zykova, T.A., Vinokurov, L.M., Markova, L.F., Kozlovskaya, E.P., and Elyakov, G.B., Bioorg. Khim., 1985, vol. 11, pp. 293–301.

    CAS  Google Scholar 

  15. Ishida, M., Minagawa, S., Miyauchi, K., Shimakura, K., Nagashima, Y., and Shiomi, K., Fisheries Sci., 1997, vol. 63, pp. 794–798.

    CAS  Google Scholar 

  16. Minagawa, S., Ishida, M., Shimakura, K., Nagashima, Y., and Shiomi, K., Comp. Biochem. Physiol. B: Biochem. Mol. Biol., 1997, vol. 118, pp. 381–386.

    Article  CAS  Google Scholar 

  17. Mebs, D. and Gebauer, E., Toxicon, 1980, vol. 18, pp. 257–264.

    Google Scholar 

  18. Minagawa, S., Sugiyama, M., Isida, M., Nagashima, Y., and Shiomi, K., Comp. Biochem. Physiol., 2008, vol. 150, pp. 240–245.

    Article  Google Scholar 

  19. Honma, T., Kawahata, S., Ishida, M., Nagai, H., Nagashima, Y., and Shiomi, K., Peptides, 2008, vol. 29, pp. 536–544.

    Article  PubMed  CAS  Google Scholar 

  20. Schweitz, H., Bruhn, T., Guillemar, E., Moinier, D., Lancelin, J.M., Béress, L., and Lazdunski, M.J., Biol. Chem., 1995, vol. 270, pp. 25121–25126.

    Article  CAS  Google Scholar 

  21. Sokotun, I.N., Il’ina, A.P., Monastyrnaya, M.M., Leichenko, E.V., Es’kov, A.A., Anastyuk, S.D., and Kozlovskaya, E.P., Biochemistry (Moscow), 2007, vol. 72, pp. 301–306.

    Article  CAS  Google Scholar 

  22. Andreev, Y.A., Kozlov, S.A., Koshelev, S.G., Ivanova, E.A., Monastyrnaya, M.M., Kozlovskaya, E.P., and Grishin, E.V., J. Biol. Chem., 2008, vol. 283, pp. 23914–23921.

    Article  PubMed  CAS  Google Scholar 

  23. Kozlov, S.A., Andreev, Ya.A., Murashev, A.N., Skobtsov, D.I., D’yachenko, I.A., and Grishin, E.V., Russ. J. Bioorg. Chem., 2009, vol. 35, pp. 789–798.

    Article  CAS  Google Scholar 

  24. Andreev, Ya.A., Kozlov, S.A., Kozlovskaya, E.P., and Grishin, E.V., Dokl. Akad. Nauk, 2009, vol. 424, pp. 46–48.

    CAS  Google Scholar 

  25. Montell, C., Sci. STKE, 2005, vol. 272, re3. DOI: 10.1126/stke.2722005re3.

  26. Moiseenkova-Bell, V.Y. and Wensel, T.G., J. Gen. Physiol., 2009, vol. 133, pp. 239–244.

    Article  PubMed  CAS  Google Scholar 

  27. Gaudet, R., J. Physiol., 2008, vol. 586, pp. 3565–3575.

    Article  PubMed  CAS  Google Scholar 

  28. Veronesi, B. and Oortgiesen, M., Toxicol. Sci., 2006, vol. 89, pp. 1–3.

    Article  PubMed  CAS  Google Scholar 

  29. Caterina, M.J., Leffler, A., Malmberg, A.B., Martin, W.J., Trafton, J., Petersen-Zeitz, K.R., Koltzenburg, M., Basbaum, A.I., and Julius, D., Science, 2000, vol. 288, pp. 306–313.

    Article  PubMed  CAS  Google Scholar 

  30. Fernández-Ballester, G. and Ferrer-Montiel, A., J. Membr. Biol., 2008, vol. 223, pp. 161–172.

    Article  PubMed  Google Scholar 

  31. Rosenbaum, T., Nat. Struct. Mol. Biol., 2009, vol. 16, pp. 704–710.

    Article  PubMed  Google Scholar 

  32. Kym, P.R., Kort, M.E., and Hutchins, C.W., Biochem. Pharmacol., 2009, vol. 78, pp. 211–216.

    Article  PubMed  CAS  Google Scholar 

  33. Roberts, L.A. and Connor, M., Recent Pat. CNS Drug Discov., 2006, vol. 1, pp. 65–76.

    Article  PubMed  CAS  Google Scholar 

  34. Szallasi, A., Cortright, D.N., Blum, C.A., and Eid, S.R., Nat. Rev. Drug. Discov., 2007, vol. 6, pp. 357–372.

    Article  PubMed  CAS  Google Scholar 

  35. Immke, D.C. and Gavva, N.R., Semin. Cell Dev. Biol., 2006, vol. 17, pp. 582–591.

    Article  PubMed  CAS  Google Scholar 

  36. Antuch, W., Berndt, D.K., Chavez, A.M., Delfin, J., and Wuthrich, K., Eur. J. Biochem., 1993, vol. 212, pp. 675–684.

    Article  PubMed  CAS  Google Scholar 

  37. Guex, N. and Peitsch, M.C., Electrophores, 1997, vol. 18, pp. 2714–2723.

    Article  CAS  Google Scholar 

  38. Peitsch, M.C., Nat. Biotechnol., 1995, vol. 13, pp. 658–660.

    Article  CAS  Google Scholar 

  39. Arnold, K., Bordoli, L., Kopp, J., and Schwede, T., Bioinformatics, 2006, vol. 22, pp. 195–201.

    Article  PubMed  CAS  Google Scholar 

  40. Kiefer, F., Arnold, K., Kunzli, M., Bordoli, L., and Schwede, T., Nucleic Acids Res., 2009, vol. 37, pp. 387–392.

    Article  Google Scholar 

  41. Laskowski, R.A., MacArthur, M.W., Moss, D.S., and Thornton, J.M., J. App. Cryst., 1993, vol. 26, pp. 283–291.

    Article  CAS  Google Scholar 

  42. Laskowski, R.A., Rullman, J.A., MacArthur, M.W., Kaptein, R., and Thornton, J.M., J. Biomol. NMR, 1996, vol. 8, pp. 477–486.

    Article  PubMed  CAS  Google Scholar 

  43. Koradi, R., Billeter, M., and Wüthrich, K., J. Mol. Grap., 1996, vol. 14, pp. 51–55.

    Article  CAS  Google Scholar 

  44. Kawamura, K., Yamada, T., Kurihara, K., Tamada, T., Kuroki, R., Tanaka, I., Takahashi, H., and Niimura, N., Acta Crystallogr., 2011, vol. 67, pp. 140–148.

    Google Scholar 

  45. Vakorina, T.I., Gladkikh, I.N., Monastyrnaya, M.M., and Kozlovskaya, E.P., Russ. J. Bioorg. Chem., 2011, vol. 37, pp. 277–284.

    Article  CAS  Google Scholar 

  46. Richter, S., Wenzel, A., Stein, M., Gabdoulline, R.R., and Wade, R.C., Nucl. Acids Res., 2008, vol. 36, pp. 276–280.

    Article  Google Scholar 

  47. De Rienzo, F., Gabdoulline, R.R., Menziani, M.C., De Benedetti, P.G., and Wade, R.C., Biophys. J., 2001, vol. 81, pp. 3090–3104.

    Article  PubMed  Google Scholar 

  48. Holtje, H.-D., Sippl, W., Rognan, D., and Folkers, G., Molecular Modeling. Basic Principles and Applications, Weinheim: WILEY-VCH Verlag GmbH Co. KGaA, 2008.

    Google Scholar 

  49. Caterina, M.J., Schumacher, M.A., Tominaga, M., Rosen, T.A., Levine, J.D., and Julius, D., Nature, 1997, vol. 389, pp. 816–824.

    Article  PubMed  CAS  Google Scholar 

  50. Tominaga, M. and Tominaga, T., Pflugers Arc., 2005, vol. 451, pp. 143–150.

    Article  CAS  Google Scholar 

  51. Brauchi, S., Orta, G., Mascayano, C., Salazar, M., Raddatz, N., Urbina, H., Rosenmann, E., Gonzalez-Nilo, F., and Latorre, R., Proc. Natl. Acad. Sci. U. S. A., 2007, vol. 104, pp. 10246–10251.

    Article  PubMed  CAS  Google Scholar 

  52. Lishko, P.V., Procko, E., Jin, X., Phelps, C.B., and Gaudet, R., Neuron, 2007, vol. 54, pp. 905–918.

    Article  PubMed  CAS  Google Scholar 

  53. Numazaki, M., Tominaga, T., Takeuchi, K., Murayama, N., Toyooka, H., and Tominaga, M., Proc. Natl. Acad. Sci. U. S. A., 2003, vol. 100, pp. 8002–8006.

    Article  PubMed  CAS  Google Scholar 

  54. Jordt, S.E. and Julius, D., Cell, 2002, vol. 108, pp. 421–430.

    Article  PubMed  CAS  Google Scholar 

  55. Lazar, J., Braun, D.C., Tóth, A., Wang, Y., Pearce, L.V., Pavlyukovets, V.A., Blumberg, P.M., Garfield, S.H., Wincovitch, S., Choi, H.-K., and Lee, J., Mol. Pharmacol., 2006, vol. 69, pp. 1166–1173.

    Article  PubMed  CAS  Google Scholar 

  56. Zelepuga, E.A., Ponomareva, R.B., Kolikov, V.M., Pautov, V.D., Sheveleva, T.V., Medvedev, M.L., Tret’yak, T.M., and Terpelovskaya, O.N., Biomed. Khim., 2003, no. 6, pp. 588–596.

  57. Terpelovskaya, O.N., Zelepuga, E.A., Ponomareva, R.B., and Tret’yak, T.M., Vopr. Med. Khim., 1993, vol. 39, no. 5, pp. 41–43.

    Google Scholar 

  58. Lange, A., Giller, K., Hornig, S., Martin-Eauclaire, M.-F., Pongs, O., Becker, S., and Baldus, M., Nature, 2006, vol. 440, pp. 959–962.

    Article  PubMed  CAS  Google Scholar 

  59. Janin, J., Bahadur, R.P., and Chakrabarti, P., Q. Rev. Biophys., 2008, vol. 41, pp. 133–180.

    Article  PubMed  CAS  Google Scholar 

  60. http://mouse.belozersky.msu.ru/npidb/cgi-bin/hftri.pl.

  61. Krowarsch, D., Zakrzewska, M., Smalas, A.O., and Otlewski, J., Protein Pept. Lett., 2005, vol. 12, pp. 403–407.

    Article  PubMed  CAS  Google Scholar 

  62. Ashcroft, F., Benos, D., Bezanilla, F., Chien, K., Choe, S., Clapham, D., Dougherty, D., Lazdunski, M., Levitan, I., Lewis, R., Liss, B., Lummis, S., Miller, C., North, A., Peters, J., Sanguinetti, M., Smart, T., Stuhmer, W., Vandenberg, J., and Verkman, A., Nat. Rev. Drug. Discov., vol. 3, pp. 237–278.

  63. Rosenbaum, T. and Simon, S.A., TRPV1 Receptors and Signal Transduction in TRP Ion Channel Function in Sensory Transduction and Cellular Signaling Cascades, Heller, S. and Liedtke, W.B., Eds., CRC Press, 2007, pp. 69–84.

  64. Il’ina, A., Lipkin, A., Barsova, E., Issaeva, M., Leychenko, E., Guzev, K., Monastyrnaya, M., Lukyanov, S., and Kozlovskaya, E., Toxicon, 2006, vol. 47, pp. 517–520.

    Article  PubMed  Google Scholar 

  65. Kumar, S., Dudley, J., Nei, M., and Tamura, K., Brief. Bioinform., 2008, vol. 9, pp. 299–306.

    Article  PubMed  CAS  Google Scholar 

  66. Altschul, S.F., Gish, W., Miller, W., Myers, E.W., and Lipman, D.J., J. Mol. Biol., 1990, vol. 215, pp. 403–410.

    PubMed  CAS  Google Scholar 

  67. Schneidman-Duhovny, D., Inbar, R., Nussinov, Y., and Wolfson, H.J., Nucleic Acids Res., 2005, vol. 33, pp. 363–367.

    Article  Google Scholar 

  68. Mashiach, E., Schneidman-Duhovny, D., Andrusier, N., Nussinov, R., and Wolfson, H.J., Nucleic Acids Res., 2008, vol. 36, pp. 229–232.

    Article  Google Scholar 

  69. Reynolds, C., Damerell, D., and Jones, S., Bioinformatics, 2009, vol. 25, pp. 413–414.

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Pacific Institute of Bioorganic Chemistry, Far East Branch, Russian Academy of Sciences, pr. 100 let Vladivostoku 159, Vladivostok, 690022, Russia

    E. A. Zelepuga, V. M. Tabakmakher, V. E. Chausova, M. M. Monastyrnaya, M. P. Isaeva & E. P. Kozlovskaya

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  1. E. A. Zelepuga
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  2. V. M. Tabakmakher
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  3. V. E. Chausova
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  4. M. M. Monastyrnaya
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  5. M. P. Isaeva
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  6. E. P. Kozlovskaya
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Correspondence to E. A. Zelepuga or V. M. Tabakmakher.

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Original Russian Text © E.A. Zelepuga, V.M. Tabakmakher, V.E. Chausova, M.M. Monastyrnaya, M.P. Isaeva, E.P. Kozlovskaya, 2012, published in Bioorganicheskaya Khimiya, 2012, Vol. 38, No. 2, pp. 185–198.

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Zelepuga, E.A., Tabakmakher, V.M., Chausova, V.E. et al. Interaction of sea anemone Heteractis crispa Kunitz type polypeptides with pain vanilloid receptor TRPV1: In silico investigation. Russ J Bioorg Chem 38, 159–170 (2012). https://doi.org/10.1134/S106816201202015X

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