Applied Microbiology and Biotechnology

, Volume 62, Issue 5–6, pp 517–522 | Cite as

Vesicular transport route of horseradish C1a peroxidase is regulated by N- and C-terminal propeptides in tobacco cells

Original Paper

Abstract.

Peroxidases (PRX, EC 1.11.1.7) are widely distributed across microorganisms, plants, and animals; and, in plants, they have been implicated in a variety of secondary metabolic reactions. In particular, horseradish (Armoracia rusticana) root represents the main source of commercial PRX production. The prxC1a gene, which encodes horseradish PRX (HRP) C, is expressed mainly in the roots and stems of the horseradish plant. HRP C1a protein is shown to be synthesized as a preprotein with both a N-terminal (NTPP) and a C-terminal propeptide (CTPP). These propeptides, which might be responsible for intracellular localization or secretion, are removed before or concomitant with production of the mature protein. We investigated the functional role of HRP C1a NTPP and CTPP in the determination of the vesicular transport route, using an analytical system of transgenically cultured tobacco cells (Nicotiana tabacum, BY2). Here, we report that NTPP and CTPP are necessary and sufficient for accurate localization of mature HRP C1a protein to vacuoles of the vesicular transport system. We also demonstrate that HRP C1a derived from a preprotein lacking CTPP is shunted into the secretory pathway.

Keywords

Enhance Green Fluorescence Protein Vesicular Transport Enhance Green Fluorescence Protein Gene Enhance Green Fluorescence Protein Fluorescence Sporamin 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgements.

We are grateful to Dr. Kazuhito Fujiyama of Osaka University for useful discussion and for technical advice regarding the IEF experiment. We would also like to thank Dr. Masami Sekine and Dr. Ko Kato of the Nara Institute of Science and Technology for their helpful discussion. This research was supported by a Grant-in-Aid for scientific research from the Ministry of Education, Science and Culture of Japan.

References

  1. Bednarek SY, Wilkins TA, Dombrowski JE, Raikhel NV (1990) A carboxy-terminal propeptide is necessary for proper sorting of barley lectin to vacuoles of tobacco. Plant Cell 2:1145–1155Google Scholar
  2. Dombrowski JE, Schroeder MR, Bednarek SY, Raikhel NV (1993) Determination of the functional elements within the vacuolar targeting signal of barley lectin. Plant Cell 5:587–596Google Scholar
  3. Fujiyama K, Takemura H, Shibayama S, Kobayashi K, Choi J-K, Shinmyo A, Takano M, Yamada Y, Okada H (1988) Structure of the horseradish peroxidase isozyme C genes. Eur J Biochem 173:681–687PubMedGoogle Scholar
  4. Fujiyama K, Takemura H, Shinmyo A, Okada H, Takano M (1990) Genomic DNA structure of two new horseradish-peroxidase-encoding genes. Gene 89:163–169PubMedGoogle Scholar
  5. Fujiyama K, Palacpac NQ, Sakai H, Kimura Y, Shinmyo A, Yoshida T, Seki T (2001) In vivo conversion of a glycan to human compatible type by transformed tobacco cells. Biochem Biophys Res Commun 289:553–557CrossRefPubMedGoogle Scholar
  6. Gad G, Sengupta-Gopalan C, Aldo C (1998) The endoplasmic reticulum of plant cells and its role in protein maturation and iogenesis of oil bodies. Plant Mol Biol 38:1–29CrossRefPubMedGoogle Scholar
  7. Helenius A, Aebi M (2001) Intracellular functions of N-linked glycans. Science 291:2364–2369PubMedGoogle Scholar
  8. Hiraga S, Sasaki K, Ito H, Ohashi Y, Matsui H (2001) A large family of class III plant peroxidases. Plant Cell Physiol 42:462–468CrossRefPubMedGoogle Scholar
  9. Holwerda BC, Padgett HS, Rogers JC (1992) Proaleurain vacuolar targeting is mediated by short contiguous peptide interactions. Plant Cell 4:307–318Google Scholar
  10. Kawasaki T, Henmi K, Ono E, Hatakeyama S, Iwano M, Satoh H, Shimamoto K (1999) The small GTP-binding protein Rac is a regulator of cell death in plants. Proc Natl Acad Sci USA 96:10922–10926CrossRefPubMedGoogle Scholar
  11. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685PubMedGoogle Scholar
  12. Lerouge P, Cabanes-Macheteau M, Rayon C, Fischette-Laine AC, Gomord V, Faye L (1998) N-Glycoprotein biosynthesis in plants: recent developments and future trends. Plant Mol Biol 38:31–48CrossRefPubMedGoogle Scholar
  13. Matsuoka K, Nakamura K (1991) Propeptide of a precursor to a plant vacuolar protein required for vacuolar targeting. Proc Natl Acad Sci USA 88:834–838PubMedGoogle Scholar
  14. Melchers LS, Sela-Buurlage MB, Vloemans SA, Woloshuk CP, Roekel JSCV, Pen J, Elzen PJM, Cornelissen BJC (1993) Extracellular targetting of the vacuolar tobacco proteins AP24, chitinase and 1,3-glucanase in transgenic plants. Plant Mol Biol 21:583–593PubMedGoogle Scholar
  15. Muntze K (1998) Deposition of storage proteins. Plant Mol Biol 38:77–99PubMedGoogle Scholar
  16. Nakayama H, Yoshida K, Ono H, Murooka Y, Shinmyo A (2000) Ectoine, the compatible solute of Halomonas elongata, confers hyperosmotic tolerance in cultured tobacco cells. Plant Physiol 122:1239–1247CrossRefPubMedGoogle Scholar
  17. Neuhaus JM, Rogers JC (1998) Sorting of proteins to vacuoles in plant cells. Plant Mol Biol 38:127–144PubMedGoogle Scholar
  18. Neuhaus JM, Sticher L, Meins F Jr, Boller T (1991) A short C-terminal sequence is necessary and sufficient for the targeting of chitinases to the plant vacuole. Proc Natl Acad Sci USA 88:10362–10366PubMedGoogle Scholar
  19. Saalbach G, Jung R, Kunze G, Saalbach I, Adler K, Muntz K (1991) Different legumin protein domains act as vacuolar targeting signals. Plant Cell 3:695–708Google Scholar
  20. Valls LA, Hunter CP, Rothman JH, Stevens TH (1987) Protein sorting in yeast: the localization determinant of yeast vacuolar carboxypeptidase Y resides in the propeptide. Cell 48:887–897PubMedGoogle Scholar
  21. Valls LA, Winther JR, Stevens TH (1990) Yeast Carboxypeptidase Y vacuolar targeting signal is defined by four propeptide amino acids. J Cell Biol 111:361–368Google Scholar
  22. Welinder KG (1979) Amino acid sequence studies of horseradish peroxidase. Amino and carboxyl termini, cyanogen bromide and tryptic fragments, the complete sequence, and some structural characteristics of horseradish peroxidase C. Eur J Biochem 96:483–502PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • T. Matsui
    • 1
  • H. Nakayama
    • 1
  • K. Yoshida
    • 1
  • A. Shinmyo
    • 1
  1. 1.Graduate School of Biological SciencesNara Institute of Science and TechnologyNaraJapan

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