Potato Research

, Volume 39, Issue 4, pp 493–505 | Cite as

A Kunitz-type proteinase inhibitor with differential solubility during the development of potato (Solanum tuberosum L.) tubers

  • Cristina Mitsumori
  • K. Yamagishi
  • T. Itoh
  • Y. Kikuta
Article
  • 24 Downloads

Summary

Two potato (Solanum tuberosum L., cv. Irish Cobbler) Kunitz-type proteinase inhibitors (PKPI) were previously described to be present in the soluble fraction of proteins from tubers in the early stages of development. One of them became insoluble in mature tubers, being extractable from this material in presence of urea. Amino acid sequencing showed that the soluble and insoluble PKPI were identical to each other. Also, immunolocalization using the protein A-gold method showed that both proteins were present inside the vacuole in free (intravacuolar space) and aggregated forms. The density of PKPI in the vacuolar protein aggregates increased from developing to mature tubers. showing that the soluble-insoluble state of this protein is related to the aggregation levels. Purified PKPI precipitated in vitro. mainly in presence of high calcium concentrations and low pH, but this precipitated form was not as stable as aggregates found in vivo. Based on the results obtained, a model of PKPI insolubilization in vivo is discussed.

Additional keywords

immunocytology protein solubility 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Ashikari, Y., Y. Arata & H. Hamaguchi, 1985. pH-induced unfolding of the constant fragment of the immunoglobulin light chain: effect of the reduction of the intrachain disulfide bond.Journal of Biochemistry 97: 517–528.PubMedGoogle Scholar
  2. Bagga, S., H. Adams, J.D. Kem & C. Sengupta-Gopalan, 1995. Accumulation of 15-kilodalton zein in novel protein bodies in transgenic tobacco.Plant Physiology 107: 13–23.PubMedGoogle Scholar
  3. Bishop, P., F. Pearce, J.E. Bryant & C.A. Ryan, 1984. Isolation and characterization of the proteinase inhibitor inducing factor from tomato leaves: identity and activity of poly- and oligogalacturonide fragments.Journal of Biological Chemistry 259: 13172–13177.PubMedGoogle Scholar
  4. Chrispeels, M.J. & N.V. Raikhel, 1992. Short peptide domains target proteins to plant vacuoles.Cell 68: 613–616.PubMedCrossRefGoogle Scholar
  5. Cleland, J.L. & I.C.D. Wang, 1990. Refolding and aggregation of bovine carbonic anhydrase B: quasi-elastic light scattering analysis.Biochemistry 29: 11072–11078.PubMedGoogle Scholar
  6. De Young, L.R., K.A. Dill & A.L. Fink, 1993. Aggregation and denaturation of apomyoglobin in aqueous urea solutions.Biochemistry 32: 3877–3886.PubMedGoogle Scholar
  7. Farmer, E.E. & C.A. Ryan, 1992. Octadecanoid precursors of jasmonic acid activate the synthesis of wound-inducible proteinase inhibitors.Plant Cell 4: 129–134.PubMedCrossRefGoogle Scholar
  8. Goldberg, M.E., R. Rudolph & R. Jaenicke, 1991. A kinetic study of the competition between renaturation and aggregation during the refolding of denatured-reduced egg white lysozyme.Biochemistry 30: 2790–2797.PubMedGoogle Scholar
  9. Hawke, D. & P. Yuan, 1987. S-pyridylethylation of cystine residues.User Bulletin, model n 470A/477A, n°28. Applied Biosystems.Google Scholar
  10. Hemsley, A.R., M.E. Collinson, W.L. Kovach, B. Vincent & T. Williams, 1994. The role of self-assembly in biological systems: evidence from iridescent colloidal sporopollenin inSelaginella megaspore walls.Philosophical Transactions of the Royal Society of London. Ser. B 345: 163–173.Google Scholar
  11. Hendricks, T., D. Vreugdenhil & W.J. Stiekema, 1991. Patatin and four serine proteinase inhibitor genes are differentially expressed during potato tuber development.Plant Molecular Biology 17: 385–394.Google Scholar
  12. Herde, O., H. Fuss, H. Peña-Cortés & J. Fisahn, 1995. Proteinase inhibitor II gene expression induced by electrical stimulation and control of photosynthetic activity in tomato plants.Plant Cell Physiology 36: 737–742.Google Scholar
  13. Hirano, H., 1988. Microsequence analysis of proteins electroblotted from polyacrylamide gels.Proteins, Nucleic Acids and Enzymes (Jap) 33: 2388–2396.Google Scholar
  14. Horisberger, M., 1981. Colloidal gold: a cytochemical marker for light and fluorescent microscopy and for transmission and scanning electron microscopy. In: O. Johari (Ed.). Scanning Electron Microscopy, vol 2. SEM Inc., AMF O'Hare Chicago, pp 9/31.Google Scholar
  15. Ishikawa, A., S. Ohta, K. Matsuoka, T. Hattori & K. Nakamura, 1994. A family of potato genes that encode Kunitz-type proteinase inhibitors: structural comparisons and differential expression.Plant Cell Physiology 35: 303–312.PubMedGoogle Scholar
  16. Kwon, K.-S., S. Lee & M.-H. Yu, 1995. Refolding of α1-antitrypsin expressed as inclusion bodies inEscherichia coli: characterization of aggregation.Biochimica et Biophysica Acta 1247: 179–184.PubMedGoogle Scholar
  17. Laemmli, U.K., 1970. Cleavage of structural proteins during the assembly of head of bacteriophage T4.Nature 227: 680–685.PubMedCrossRefGoogle Scholar
  18. Mitsumori, C., K. Yamagishi, K. Fujino & Y. Kikuta, 1994. Detection of immunologically related Kunitz and Bowman-Birk proteinase inhibitors expressed during potato development.Plant Molecular Biology 26: 961–969.PubMedCrossRefGoogle Scholar
  19. Nishimura, M., 1987. Microbodies.Proteins. Nucleic Acids and Enzymes (Jap) 30: S-37–42.Google Scholar
  20. Ono, T., S. Katho & K. Mothizuki, 1993. Influences of calcium and pH on protein solubility in soybean milk.Bioscience, Biotechnology and Biochemistry 57: 24–28.Google Scholar
  21. Pelham, H.R.B., 1989. Control of proteins exit from the endoplasmic reticulum.Annual Review of Cell Biology 5: 1–23.PubMedCrossRefGoogle Scholar
  22. Peña-Cortés, H., T. Albrecht, S. Prat, E.W. Weiler & L. Willmitzer, 1993. Aspirin prevents wound-induced gene expression in tomato leaves by blocking jasmonic acid biosynthesis.Planta 191: 123–128.Google Scholar
  23. Ryan, C.A. & G. An, 1988. Molecular biology of wound-inducible proteinase inhibitors in plants.Plant Cell Environment 11: 345–349.Google Scholar
  24. Suh, S.G., J.E. Peterson, W.J. Stiekema & D.J. Hannapel, 1990. Purification and characterization of the 22-Kilodalton potato tuber protein.Plant Physiology 94: 40–45.Google Scholar
  25. Vitale, A. & M.J. Chrispeels, 1992. Sorting of proteins to the vacuoles of plant cells.BioEssarys 14: 151–160.Google Scholar
  26. Walsh, T.A. & W.P. Twitchell, 1991. Two Kunitz-type proteinase inhibitors from potato tubers.Plant Physiology 97: 15–18.Google Scholar
  27. Yamagishi, K., C. Mitsumori, Y. Kikuta, 1991. Nucleotide sequence of a cDNA encoding the putative trypsin inhibitor in potato tubers.Plant Molecular Biology 17: 287–288.PubMedCrossRefGoogle Scholar
  28. Yamagishi, K., C. Mitsumori, K. Takahashi, K. Fujino, Y. Koda & Y. Kikuta, 1993. Jasmonic acid-inducible gene expression of a Kunitz-type proteinase inhibitor in potato tuber disks.Plant Molecular Biology 21: 539–41.PubMedCrossRefGoogle Scholar
  29. Yokota, S., 1991. Practice in the postembedding immunocytochemical techniques.The Cell (Tokyo) 23: 425–429.Google Scholar

Copyright information

© Kluwer Academic Publishers 1996

Authors and Affiliations

  • Cristina Mitsumori
    • 1
  • K. Yamagishi
    • 1
  • T. Itoh
    • 2
  • Y. Kikuta
    • 1
  1. 1.Laboratory of Crop Physiology, Faculty of AgricultureHokkaido UniversitySapporoJapan
  2. 2.Laboratory of Electronic Microscopy. Faculty of AgricultureHokkaido UniversitySapporoJapan

Personalised recommendations