Eur. J. Entomol. 108 (4): 711-715, 2011 | DOI: 10.14411/eje.2011.091

Contribution to the biochemical characterization of the silk and structure characterization of the cocoons of the horse chestnut leaf miner Cameraria ohridella (Lepidoptera: Gracillariidae)

Veronika HNĚVSOVÁ1, Dalibor KODRÍK*,2,3, František WEYDA2,3
1 Gymnasium, Jírovcova 8, 371 61 České Budějovice, Czech Republic
2 Institute of Entomology, Biology Centre, Academy of Sciences and
3 Faculty of Science, University of South Bohemia, Branišovská 31, 370 05 České Budějovice, Czech Republic

This study of the proteins in the silk of the summer and winter cocoons of the horse chestnut leaf miner Cameraria ohridella revealed they can inhibit protease activity. The inhibitory activity of the summer silk was higher against both the fungal proteinase K and bacterial subtilisin than that of the winter silk. Also, the winter silk was more effective in inhibiting proteinase K than subtilisin. Further, it was demonstrated that some of the silk proteins are glycosylated probably by mannose carbohydrates. An electron microscopy study of the cocoons revealed the presence of silk fibres with different diameters in some pupal chambers.

Keywords: Silk, cocoon, pupal chamber, tissue of leaf mine, Cameraria ohridella, Gracillariidae, protease inhibitor, glycoprotein, electron microscopy

Received: April 28, 2011; Revised: July 26, 2011; Accepted: July 26, 2011; Published: October 3, 2011  Show citation

ACS AIP APA ASA Harvard Chicago IEEE ISO690 MLA NLM Turabian Vancouver
HNĚVSOVÁ, V., KODRÍK, D., & WEYDA, F. (2011). Contribution to the biochemical characterization of the silk and structure characterization of the cocoons of the horse chestnut leaf miner Cameraria ohridella (Lepidoptera: Gracillariidae). EJE108(4), 711-715. doi: 10.14411/eje.2011.091
Download citation

References

  1. CHOU J. & CHOU T.C. 1987: Dose-Effect Analysis with Microcomputers. Biosoft, Cambridge, UK, 93 pp
  2. CONNOR E.F. & TAVERNER M.P. 1997: The evolution and adaptive significance of the leaf-mining habit. Oikos 79: 6-25 Go to original source...
  3. COUBLE P., MOINE A., GAREL A. & PRUDHOMME J.C. 1983: Developmental variations of a nonfibroin mRNA of Bombyx mori silkgland, encoding for a low-molecular-weight silk protein. Dev. Biol. 97: 398-407 Go to original source...
  4. FEDIC R., ZUROVEC M. & SEHNAL F. 2002: The silk of Lepidoptera. J. Insect Biotech. Sericol. 71: 1-15
  5. GAMO T., INOUCHI T. & LAUFER H. 1977: Polypeptides of fibroin and sericin secreted from the different sections of the silk gland in Bombyx mori. Insect Biochem. 7: 285-295 Go to original source...
  6. GRUBHOFFER L., GUIRAKHOO F., HEINZ F.X. & KUNZ C. 1990: Interaction of tick-borne encephalitis virus protein E with labelled lectins. In Kocourek J. & Freed. D.L. (eds): Lectins: Biology, Biochemistry, Clinical Biochemistry. Vol. 7. Sigma Chemical Company, Saint Louis, pp. 313-319
  7. GRZELAK K., COUBLE P., GAREL A. & KLUDKIEWICZ B. 1988: Low molecular weight silk proteins in Galleria mellonella. Insect Biochem. 18: 223-228 Go to original source...
  8. INOUE S., TANAKA K., ARISAKA F., KIMURA S. OHMOTO K. & MIZUNO S. 2000: Silk fibroin of Bombyx mori is secreted, assembling a high molecular mass elementary unit consisting of H-chain, L-chain, and P-25, with a 6:6:1 molar ratio. J. Biol. Chem. 275: 40517-40528 Go to original source...
  9. KIMURA K., OYAMA F., UEDA H., MIZUNO S. & SHIMURA K. 1985: Molecular cloning of the fibroin light chain complementary DNA and its use in the study of the expression of the light chain gene in the posterior silk gland of Bombyx mori. Experientia 41: 1167-1171 Go to original source...
  10. KIRKEBY S., MOE D. & BOG-HANSEN T.C. 1993: The silver staining procedure of sodium dodecyl sulphate-gels may be accelerated by shortening fixation time. Electrophoresis 14: 51-55. Go to original source...
  11. KLUDKIEWICZ B., KODRIK D., GRZELAK K., XAVIER N. & SEHNAL F. 2005: Structurally unique recombinant Kazal-type proteinase inhibitor retains activity when terminally extended and glycosylated. Protein Expres. Purif. 43: 94-102 Go to original source...
  12. KODRIK D. 1992: Small protein components of the cocoons in Galleria mellonella (Lepidoptera, Pyralidae) and Bombyx mori (Lepidoptera, Bombycidae). Acta Entomol. Bohemoslov. 89: 269-274
  13. KODRIK D. & SEHNAL F. 1994: Juvenile hormone counteracts the action of ecdysterone on silk glands of Galleria mellonella L. (Lepidoptera: Pyralidae). Int. J. Insect Morphol. Embryol. 23: 39-56 Go to original source...
  14. LAEMMLI U.K. 1970: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685 Go to original source...
  15. NIRMALA X., KODRIK D., ZUROVEC M. & SEHNAL F. 2001a: Insect silk contains both a Kunitz-type and a unique Kazal-type proteinase inhibitors. Eur. J. Biochem. 268: 2064-2073 Go to original source...
  16. NIRMALA X., MITA K., VANISREE V., ZUROVEC M. & SEHNAL F. 2001b: Identification of four small molecular mass proteins in the silk of Bombyx mori. Insect Mol. Biol. 10: 437-445 Go to original source...
  17. SASAKI T. & NODA H. 1973: Studies on silk fibroin of Bombyx mori directly extracted from the silk gland. I. Molecular weight determination in guanidine hydrochloride or urea solutions. Biochim. Biophys. Acta 310: 76-90 Go to original source...
  18. SEHNAL F. & AKAI H. 1990: Insects silk glands: their types, development and function, and effects of environmental factors and morphogenetic hormones on them. Int. J. Insect Morphol. Embryol. 19: 79-132 Go to original source...
  19. SEHNAL F. & ZUROVEC M. 2004: Construction of silk fiber core in Lepidoptera. Biomacromolecules 5: 666-674 Go to original source...
  20. SKUHRAVY V. 1998: On the leaf mining moth Cameraria ohridella Desch. & Dim. (Lep., Lithocolletidae) attacking Aesculus hippocastanum L. in the Czech Republic. Anz. Schadlingsk. Pflanzensch. Umweltsch. 71: 81-84 Go to original source...
  21. STOSCHECK C.M. 1990: Quantitation of proteins. Methods Enzymol. 182: 50-68 Go to original source...
  22. TAKEI F., KIKUCHI Y., KIKUCHI A., MIZUNO S. & SHIMURA K. 1987: Further evidence for importance of the subunit combination of silk fibroin in its efficient secretion from the posterior silk gland cells. J. Cell Biol. 105: 175-180 Go to original source...
  23. TANAKA K., INOUE S. & MIZUNO S. 1999: Hydrophobic interaction of P25, containing Asn-linked oligosaccharide chains, with the H-L complex of silk fibroin produced by Bombyx mori. Insect Biochem. Mol. Biol. 29: 269-276 Go to original source...
  24. TOKUTAKE S. 1980: Isolation of the smallest component of silk protein. Biochem. J. 187: 413-417 Go to original source...
  25. TOWBIN H., STAEHLIN T. & GORDON J. 1979: Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc. Natl. Acad. Sci. U. S. A. 76: 4350-4354 Go to original source...
  26. ZUROVEC M., VASKOVA M., KODRIK D., SEHNAL F. & KUMARAN A.K. 1995: Light-chain fibroin of Galleria mellonella L. Mol. Gen. Genet. 247: 1-6 Go to original source...
  27. ZUROVEC M., YANG C., KODRIK D. & SEHNAL F. 1998a: Identification of a novel type of silk protein and regulation of its expression. J. Biol. Chem. 273: 15423-15428 Go to original source...
  28. ZUROVEC M., KODRIK D., YANG C., SEHNAL F. & SCHELLER K. 1998b: P25 component of Galleria silk. Mol. Gen. Genet. 257: 264-270 Go to original source...

This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (CC BY 4.0), which permits use, distribution, and reproduction in any medium, provided the original publication is properly cited. No use, distribution or reproduction is permitted which does not comply with these terms.