Hct-A Is a New Actinoporin Family from the Heteractis Crispa Sea Anemone

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Abstract

Several new actinoporin isoforms with molecular weights of 18995.5 to 19398.7 Da exhibiting a high hemolytic activity were isolated from the tropical sea anemone Heteractis crispa using a combination of liquid chromatography techniques. The actinoporins were demonstrated to occur as mono-, di-, and trimers in aqueous solutions. The sequences of the genes encoding actinoporins were identified, and the amino acid sequences of the new polypeptides belonging to the Hct-A actinoporin family were obtained. The new acinoporins differ in their isoelectric points, the number and localization of charged amino acid residues at the functionally important N-terminal fragment of the molecule, as well as in the charge of a tetrapeptide (amino acid residues 74-77) involved in an electrostatic interaction with the cytoplasmic membrane. A recombinant actinoporin, rHct-A2, with a molecular weight of 19141 Da, pI of 9.64, and hemolytic activity of 4.0 × 104 HU/mg, was obtained. The conductivity of the ion channels formed by rHct-A2 in the BLM was demonstrated to be similar to that of the native actinoporin from H. crispa. The obtained data expand knowledge on the structural and functional relationships of actinoporins and contribute to our understanding of the functioning mechanism of these molecules, which is the basis for the development of compounds with a high biomedical potential. Currently, they are considered as models for obtaining antitumor, antibacterial, and cardiac-stimulating agents.

About the authors

E. V. Leichenko

Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch of the Russian Academy of Sciences

Author for correspondence.
Email: 969844@gmail.com
Russian Federation

M. M. Monastirnaya

Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch of the Russian Academy of Sciences

Email: 969844@gmail.com
Russian Federation

E. A. Zelepuga

Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch of the Russian Academy of Sciences

Email: 969844@gmail.com
Russian Federation

E. S. Tkacheva

Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch of the Russian Academy of Sciences

Email: 969844@gmail.com
Russian Federation

M. P Isaeva

Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch of the Russian Academy of Sciences

Email: 969844@gmail.com
Russian Federation

G. N. Likhatskaya

Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch of the Russian Academy of Sciences

Email: 969844@gmail.com
Russian Federation

S. D. Anastyuk

Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch of the Russian Academy of Sciences

Email: 969844@gmail.com
Russian Federation

E. P. Kozlovskaya

Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch of the Russian Academy of Sciences

Email: 969844@gmail.com
Russian Federation

References

  1. Parker M.W., Feil S.C. // Prog. Biophys. Mol. Biol. 2005, V.88, №6202, P.91-142
  2. Chi V., Pennington M.W., Norton R.S., Tarcha E.J., Londono L.M., Sims-Fahey B., Upadhyay S.K., Lakey J.T., Iadonato S., Wulff H. // Toxicon. 2012, V.59, №4, P.529-546
  3. Frazão B., Vasconcelos V., Antunes A. // Mar. Drugs. 2012, V.10, №8, P.1812-1851
  4. Turk T.J. // Toxicol. Toxin. Rev. 1991, V.10, P.223-262
  5. Anderluh G., Maček P. // Toxicon. 2002, V.40, №2, P.111-124
  6. Tejuca M., Anderluh G., Dalla Serra M. // Toxicon. 2009, V.54, №8, P.1206-1214
  7. Fedorov S., Dyshlovoy S., Monastyrnaya M., Shubina L., Leychenko E., Kozlovskaya E., Jin J.O., Kwak J.-Y., Bode A.M., Dong Z. // Toxicon. 2010, V.55, №4, P.811-817
  8. Batista U., Maček P., Sedmak B. // Cell Biol. Int. Rep. 1990, V.14, №11, P.1013-1024
  9. Norton R.S., Bobek G., Ivanov J.O., Thomson M., Beer E.F., Mortiz R.L., Simpson R.J. // Toxicon. 1990, V.28, №1, P.29-41
  10. Migues P.V., Leal R.B., Mantovanni M., Nicolau M., Gabilan N.H. // NeiroReport. 1999, V.10, №1, P.67-70
  11. Soletti R.C., Alves T., Vernal J., Terenzi H., Anderluh G., Borges H.L., Gabilan N.H., Moura-Neto V. // Anticancer Res. 2010, V.30, №4, P.1209-1215
  12. Monastyrnaya M.M., Zykova T.A., Apalikova O.V., Shwets T.V., Kozlovskaya E.P. // Toxicon. 2002, V.40, №8, P.1197-1217
  13. Lowry O.H., Rosebrough N.J., Fearr A.L., Randall R.J. // J. Biol. Chem. 1951, V.193, №1, P.265-275
  14. Il´ina A., Lipkin A., Barsova E., Issaeva M., Leychenko E., Guzev K., Monastyrnaya M., Lukyanov S., Kozlovskaya E. // Toxicon. 2006, V.47, №5, P.517-520
  15. Sambrook J., Russel D.W. // Molecular cloning. Laboratory Manual. 3rd ed. New York: Cold Spring Harbor Laboratory Press, 2001. 2001, P.1.31-1.58
  16. Sanger F., Nicklen S., Coulson A.R. // Proc. Natl. Acad. Sci. USA. 1977, V.74, №12, P.5463-5467
  17. Laemmli U.K. // Nature 1970, V.227, №5259, P.680-685
  18. Muller P., Rudin D.O., Tien H.T., Wescott W.C. // J. Phys. Chem. 1963, V.67, P.534-535
  19. Guex N., Peitsch M.C. // Electrophoresis. 1997, V.18, №15, P.2714-2723
  20. Guex N., Peitsch M.C., Schwede T. // Electrophoresis. 2009, V.30, S1, P.162-173
  21. Mancheño J.M., Martín-Benito J., Martínez-Ripoll M., Gavilanes J.G., Hermoso J.A. // Structure. 2003, V.11, №11, P.1319-1328
  22. // url www.rcsb.org/
  23. // Molecular Operating Environment (MOE), 2013.08; Chemical Computing Group Inc. Chemical Computing Group Inc., 1010 Sherbooke St. West, Suite #910, Montreal, QC, Canada, H3A 2R7, 2014. 2014
  24. Lanio M.E., Morera V., Alvarez C., Tejuca M., Gómez T., Pazos F., Besada V., Martínez D., Huerta V., Padrón G. // Toxicon. 2001, V.39, №2-3, P.187-194
  25. Klyshko E.V., Issaeva M.P., Monastyrnaya M.M., Il´ina A.P., Guzev K.V., Vakorina T.I., Dmitrenok P.S., Zykova T.A., Kozlovskaya E.P. // Toxicon. 2004, V.44, №3, P.315-324
  26. Tkacheva E.S., Leychenko E.V., Monastyrnaya M.M., Issaeva M.P., Zelepuga E.A., Anastyuk S.D., Dmitrenok P.S., Kozlovskaya E.P. // Biochemistry. 2011, V.76, №10, P.1387-1397
  27. Alegre-Cebollada J., Cunietti M., Herrero-Galán E., Gavilanes J.G., Martínez-del-Pozo A. // J. Mol. Biol. 2008, V.382, №4, P.920-930
  28. Monastyrnaya M., Leychenko E., Issaeva M., Likhatskaya G., Zelepuga E., Kostina E., Trifonov E., Nurminski E., Kozlovskaya E. // Toxicon. 2010, V.56, №8, P.1299-1314
  29. Maček P., Lebez D. // Toxicon. 1981, V.19, №2, P.233-240
  30. Anderluh G., Pungerčar J., Štrukelj B., Maček P., Gubenšek F. // Biochem. Biophys. Res. Commun. 1996, V.220, №2, P.437-442
  31. Samejima Y., Yanagisaws M., Aoki-Tomomutsu Y., Iwasaki E., Ando J., Mebs D. // Toxicon. 2000, V.38, №2, P.259-264
  32. Nagai H., Oshiro N., Takuwa-Kuroda K., Iwanaga S., Nozaki M., Nakajima T.A. // Biosci. Biotechnol. Biochem. 2002, V.66, №12, P.2621-2625
  33. Wang Y., Yap L.L., Chua K.L., Khoo H.E. // Toxicon. 2008, V.51, №8, P.1374-1382
  34. Anderluh G., Krizaj I., Štrukelj B., Gubenšek F., Maček P., Pungerčar J. // Toxicon. 1999, V.37, №10, P.1391-1401
  35. Blumenthal K.M., Kem W.R. // J. Biol. Chem. 1983, V.258, №9, P.5574-5581
  36. Kristan K., Podlesek Z., Hojnik V., Gutierrez-Aguirre I., Gunčar G., Turk D., Gonzalez-Manas J.M., Lakey J.H., Maček P., Anderluh G. // J. Biol. Chem. 2004, V.279, №45, P.46509-46517
  37. Anderluh G., Barlič A., Potrich C., Macek P., Menestrina G. // J. Membrane Biol. 2000, V.173, №1, P.47-55
  38. Mechaly A.E., Bellomio A., Gil-Cartón D., Morante K., Valle M., González-Mañas J.M., Guérin D.M. // Structure. 2011, V.19, №2, P.181-191
  39. Bakrač B., Gutiérrez-Aguirre I., Podlesek Z., Sonnen A.F.P., Gilbert R.J.C., Macek P., Lakey J.H., Anderluh G. // J. Biol. Chem. 2009, V.283, №27, P.18665-18677
  40. Rojko N., Kristan K.C., Viero G., Zerovnik E., Maček P., Dalla Serra M., Anderluh G. // J. Biol. Chem. 2013, V.288, №33, P.23704-23715
  41. Zorec R., Tester M., Maček P., Mason W.T. // J. Membrane Biol. 1990, V.118, №3, P.243-249
  42. Meunier F.A., Frangez R., Benoit E., Ouanounou G., Rouzaire-Dubois B., Suput D., Molgo J. // Toxicon. 2000, V.38, №11, P.1547-1560
  43. Lewis R.J., Garcia M.L. // Nat. Rev. Drug Discov. 2003, V.2, №10, P.790-802
  44. Takagi J. // Biochem. Soc. Transactions. 2004, V.32, №3, P.403-406

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Copyright (c) 2014 Leichenko E.V., Monastirnaya M.M., Zelepuga E.A., Tkacheva E.S., Isaeva M.P., Likhatskaya G.N., Anastyuk S.D., Kozlovskaya E.P.

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