Biochemistry (Moscow)

, Volume 72, Issue 4, pp 430–438 | Cite as

Purification and primary structure of novel lipid transfer proteins from germinated lentil (Lens culinaris) seeds

  • E. I. Finkina
  • S. V. Balandin
  • M. V. Serebryakova
  • N. A. Potapenko
  • A. A. Tagaev
  • T. V. Ovchinnikova
Article

Abstract

A subfamily of eight novel lipid transfer proteins designated as Lc-LTP1-8 was found in the lentil Lens culinaris. Lc-LTP2, Lc-LTP4, Lc-LTP7, and Lc-LTP8 were purified from germinated lentil seeds, and their molecular masses (9268.7, 9282.7, 9121.5, 9135.5 daltons) and complete amino acid sequences were determined. The purified proteins consist of 92–93 amino acid residues, have four disulfide bonds, and inhibit growth of Agrobacterium tumefaciens. Total RNA was isolated from germinated lentil seeds, RT-PCR and cloning were performed, and the cDNAs of six LTPs were sequenced. Precursor 116–118-residue proteins with 24–25-residue signal peptides were found, and two of them are purified proteins Lc-LTP2 and Lc-LTP4.

Key words

lipid transfer proteins lentil germinated seeds primary structure antibacterial activity cDNA 

Abbreviations

FPLC

fast protein liquid chromatography

LTP

lipid transfer protein

MALDI-TOF-MS

matrix-assisted laser desorption ionization time-of-flight mass spectrometry

PR-proteins

pathogenesis-related proteins

RACE

rapid amplification of cDNA ends

RT-PCR

reverse trnascription polymerase chain reaction

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Selitrennikoff, C. P. (2001) Appl. Environ. Microbiol., 67, 2883–2894.PubMedCrossRefGoogle Scholar
  2. 2.
    Hoffmann-Sommergruber, K. (2002) Biochem. Soc. Trans., 30, 930–935.PubMedCrossRefGoogle Scholar
  3. 3.
    Castro, M. S., and Fontes, W. (2005) Prot. Pept. Lett., 12, 13–18.Google Scholar
  4. 4.
    Molina, A., Segura, A., and Garcia-Olmedo, F. (1993) FEBS Lett., 316, 119–122.PubMedCrossRefGoogle Scholar
  5. 5.
    Douliez, J., Michon, T., Elmorjani, K., and Marion, D. (2000) J. Cereal Sci., 32, 1–20.CrossRefGoogle Scholar
  6. 6.
    Kader, J. (1996) Annu. Rev. Plant Physiol. Plant Mol. Biol., 47, 627–654.PubMedCrossRefGoogle Scholar
  7. 7.
    Garcia-Olmedo, F., Molina, A., Alamillo, J. M., and Rodriguez-Palenzuella, P. (1999) Biopolymers, 47, 479–491.CrossRefGoogle Scholar
  8. 8.
    Samuel, D., Liu, Y., Cheng, C., and Lyu, P. (2002) J. Biol. Chem., 277, 35267–35273.PubMedCrossRefGoogle Scholar
  9. 9.
    Kader, J. (1997) Trends Plant Sci., 2, 66–70.CrossRefGoogle Scholar
  10. 10.
    Hughes, M. A., Dunn, M. A., Pearce, R. S., White, A. J., and Zhang, L. (1992) Plant Cell Environ., 15, 861–865.CrossRefGoogle Scholar
  11. 11.
    Hincha, D. K. (2002) Phil. Trans. R. Soc. Lond., 357, 909–916.CrossRefGoogle Scholar
  12. 12.
    Trevino, M. B., and O’Conell, M. A. (1998) Plant Physiol., 116, 1461–1468.PubMedCrossRefGoogle Scholar
  13. 13.
    Dani, V., Simon, W. J., Duranti, M., and Croy, R. R. (2005) Proteomics, 5, 737–745.PubMedCrossRefGoogle Scholar
  14. 14.
    Kristensen, A. K., Brunstedt, J., Nielsen, K. K., Roepstorff, P., and Mikkelsen, J. D. (2000) Plant Sci., 155, 31–40.PubMedCrossRefGoogle Scholar
  15. 15.
    Garcia-Olmedo, F., Molina, A., Segura, A., and Moreno, M. (1995) Trends Microbiol., 3, 72–74.PubMedCrossRefGoogle Scholar
  16. 16.
    Blein, J., Thevenot, P., Marion, D., and Ponchet, M. (2002) Trends Plant Sci., 7, 293–296.PubMedCrossRefGoogle Scholar
  17. 17.
    Park, S. Y., Jauh, G. Y., Mollet, J. C., Eckard, K. J., Nothnagel, E. A., Walling, L. L., and Lord, E. M. (2000) Plant Cell, 12, 151–163.PubMedCrossRefGoogle Scholar
  18. 18.
    Eklund, D. M., and Edqvist, J. (2003) Plant Physiol., 132, 1249–1259.PubMedCrossRefGoogle Scholar
  19. 19.
    Crimi, M., Astegno, A., Zoccatelli, G., and Esposti, M. D. (2006) Arch. Biochem. Biophys., 445, 65–71.PubMedCrossRefGoogle Scholar
  20. 20.
    Douliez, J., Pato, C., Rabesona, H., Molle, D., and Marion, D. (2001) Eur. J. Biochem., 268, 1400–1403.PubMedCrossRefGoogle Scholar
  21. 21.
    Douliez, J., Michon, T., and Marion, D. (2000) Biochim. Biophys. Acta, 1467, 65–72.CrossRefGoogle Scholar
  22. 22.
    Pato, C., Le Borgne, M., Le Baut, G., Le Pape, P., Marion, D., and Douliez, N. (2001) Biochem. Pharm., 62, 555–560.PubMedCrossRefGoogle Scholar
  23. 23.
    Shewry, P. R., Beaudoin, F., Jenkins, J., Griffiths-Jones, S., and Mills, E. N. (2002) Biochem. Soc. Trans., 30, 906–910.PubMedCrossRefGoogle Scholar
  24. 24.
    Diaz-Perales, A., Sanz, M. L., Garcia-Casado, G., Sanchez-Monge, R., Garcia-Selles, F. J., Lombardero, M., Polo, F., Gamboa, P. M., Barber, D., and Salcedo, G. (2003) J. Allergy Clin. Immunol., 111, 628–633.PubMedCrossRefGoogle Scholar
  25. 25.
    Ballmer-Weber, B. K., Wangorsch, A., Bohle, B., Kaul, S., Kundig, T., Fotisch, K., van Ree, R., and Vieths, S. (2005) Clin. Exp. Allergy, 35, 970–978.PubMedCrossRefGoogle Scholar
  26. 26.
    Schagger, H., and Jagow, G. (1987) Analyt. Biochem., 166, 368–379.PubMedCrossRefGoogle Scholar
  27. 27.
    Gray, W. R. (1967) Meth. Enzymol., 11, 139–151.CrossRefGoogle Scholar
  28. 28.
    Ovchinnikova, T., Aleshina, G., Balandin, S., Krasnosdembskaya, A., Markelov, M., Frolova, E., Leonova, Y., Tagaev, A., Krasnodembsky, E., and Kokryakov, V. (2004) FEBS Lett., 577, 209–214.PubMedCrossRefGoogle Scholar
  29. 29.
    Sambrook, J., Fritch, E. F., and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, 2nd Edn., Cold Springer Harbor Laboratory, NY.Google Scholar
  30. 30.
    Matz, M., Lukyanov, S., Bogdanova, E., Diatchenko, L., and Chenchik, A. (1999) Nucleic Acids Res., 27, 1558–1560.PubMedCrossRefGoogle Scholar
  31. 31.
    Ostergaard, J., Hojrup, P., and Knudsen, J. (1995) Biochim. Biophys. Acta, 1254, 169–179.PubMedGoogle Scholar
  32. 32.
    Soufleri, I. A., Vergnolle, C., Miginiac, E., and Kader, J. C. (1996) Planta, 199, 229–237.PubMedCrossRefGoogle Scholar
  33. 33.
    Lullien-Pellerin, V., Devaux, C., Ihorai, T., Marion, D., Pahin, V., Joudrier, P., and Gautier, M.-F. (1999) Eur. J. Biochem., 260, 861–868.PubMedCrossRefGoogle Scholar
  34. 34.
    Ge, X., Chen, J., Sun, C., and Cao, K. (2003) Prot. Eng., 16, 387–390.CrossRefGoogle Scholar

Copyright information

© Pleiades Publishing, Ltd. 2007

Authors and Affiliations

  • E. I. Finkina
    • 1
  • S. V. Balandin
    • 1
  • M. V. Serebryakova
    • 2
  • N. A. Potapenko
    • 1
  • A. A. Tagaev
    • 1
  • T. V. Ovchinnikova
    • 1
  1. 1.Shemyakin and Ovchinnikov Institute of Bioorganic ChemistryRussian Academy of SciencesMoscowRussia
  2. 2.Orekhovich Institute of Biomedical ChemistryRussian Academy of Medical SciencesMoscowRussia

Personalised recommendations