Skip to main content
Log in

Primary structure of the N-linked carbohydrate chains of Calreticulin from spinach leaves

  • Papers
  • Published:
Glycoconjugate Journal Aims and scope Submit manuscript

Abstract

Calreticulin is a multifunctional Ca2+-binding protein of the endoplasmic reticulum of most eukaryotic cells. The 56 kDa Calreticulin glycoprotein isolated from spinach (Spinacia oleracea L.) leaves was N-deglycosylated by PNGase-F digestion. The carbohydrate moiety was isolated by gel permeation chromatography and purified by high-pH anion-exchange chromatography. The fractions were investigated by 500 MHz1H-NMR spectroscopy, in combination with monosaccharide analysis and fast-atom bombardment-mass spectrometry. The following carbohydrate structure could be established as the major component (Man8GlcNAc2):

Heterogeneity was demonstrated by the presence of two minor components being Man7GlcNAc2 lacking a terminal residue (D1 or D3), compared to the major component. A cross-reactivity with an antibody against the endoplasmic reticulum retention signal HDEL was also found.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Michalak M, Milner R, Burns K, Opas M (1992)Biochem J 285: 681–92.

    PubMed  Google Scholar 

  2. Nash PD, Opas M, Michalak M (1994)Mol Cell Biochem 135: 71–8.

    PubMed  Google Scholar 

  3. Van PN, Peter F, Söling H-D (1989)J. Biol Chem 264: 17494–501.

    PubMed  Google Scholar 

  4. Peter F, Van PN, Söling H-D (1992)J Biol Chem 267: 10631–7.

    PubMed  Google Scholar 

  5. Khanna NC, Tokuda M, Waisman DM (1987)Biochem J 242: 245–51.

    PubMed  Google Scholar 

  6. Matsuoka KM, Seta K, Yamakawa Y, Okuyama T, Shinoda T, Isobe T (1994)Biochem J 298: 435–42.

    PubMed  Google Scholar 

  7. Menegazzi P, Guzzo F, Baldan B, Mariani P, Treves S (1993)Biochem Biophys Res Commun 3: 1130–5.

    Google Scholar 

  8. Hassan A-M, Wesson C, Trumble WR (1995)Biochem Biophys Res Commun 211: 54–9.

    PubMed  Google Scholar 

  9. Denecke J, Carlsson LE, Vidal S, Hoglund AS, Ek B, van Zeijl MJ, Sinjorgo KMC, Palva ET (1995)Plant Cell 7: 391–406.

    PubMed  Google Scholar 

  10. Napier RM, Trueman S, Henderson J, Boyce JM, Hawes C, Fricker MD, Venis MA (1995)J Exp Bot 46: 1603–13.

    Google Scholar 

  11. Navazio L, Baldan B, Dainese P, James P, Damiani E, Margreth A, Mariani P (1995)Plant Physiol 109: 983–90.

    PubMed  Google Scholar 

  12. Bradford MM (1976)Anal Biochem 72: 248–54.

    PubMed  Google Scholar 

  13. Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F (1956)Anal Chem 28: 350–6.

    Google Scholar 

  14. Kamerling JP, Vliegenthart JFG (1989)Carbohydrates. InClinical Biochemistry: Principles, Methods, Applications, Vol I,Mass spectrometry (Lawson AM, ed.) pp. 176–263. Berlin, New York: Walter de Gruyter.

    Google Scholar 

  15. Laemmli UK (1970)Nature 227: 680–5.

    PubMed  Google Scholar 

  16. Napier RM, Fowke LC, Hawes C, Leis M, Pelham HRB (1992)J Cell Sci 102: 261–71.

    PubMed  Google Scholar 

  17. Damm JBL, Voshol H, Hård K, Kamerling JP, Vliegenthart JFG (1989)Eur J Biochem 180: 101–10.

    PubMed  Google Scholar 

  18. Hård K, van Zadelhoff G, Moonen P, Kamerling JP, Vliegenthart JFG (1992)Eur J Biochem 209: 895–915.

    PubMed  Google Scholar 

  19. Vliegenthart JFG, Dorland L, Van Halbeek H (1983)Adv Carbohydr Chem Biochem 41: 209–374.

    Google Scholar 

  20. Neeser J-R, Del Vedovo S, Mutsaers JHGM, Vliegenthart JFG (1985)Glycoconjugate J 2: 355–64.

    Google Scholar 

  21. Hård K, Mekking A, Kamerling JP, Dacremont GAA, Vliegenthart JFG (1991)Glycoconjugate J 8: 17–28.

    Google Scholar 

  22. Van Kuik JA, Hård K, Vliegenthart JFG (1992)Carbohydrate Res 235: 53–68.

    Google Scholar 

  23. Chen F, Hayes PM, Mulrooney DM, Pan A (1994)Plant Cell 6: 835–43.

    PubMed  Google Scholar 

  24. Sturm A (1995) InGlycoproteins. New Comprehensive Biochemistry Vol. 29a (Montreuil J, Vliegenthart JFG, Schachter H, eds) pp. 521–54 Amsterdam: Elsevier.

    Google Scholar 

  25. Koch GLE (1987)J. Cell Sci 87: 491–2.

    PubMed  Google Scholar 

  26. Opas M, Dziak E, Fliegel L, Michalak M (1991)J. Cell Physiol 149: 160–71.

    PubMed  Google Scholar 

  27. Burns K, Duggan B, Atkinson EA, Famulski KS, Nemer M, Bleackley RC, Michalak M (1994)Nature 367: 476–80.

    PubMed  Google Scholar 

  28. Dedhar S, Rennie PS, Shago M, Hagesteijn C-YL, Yang H, Filmus J, Hawley RG, Bruchovsky N, Cheng H, Matusik RJ, Giguere V (1994)Nature 367: 480–3.

    PubMed  Google Scholar 

  29. White TK, Zhu Q, Tanzer ML (1995)J. Biol Chem 270: 15926–9.

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Navazio, L., Baldan, B., Mariani, P. et al. Primary structure of the N-linked carbohydrate chains of Calreticulin from spinach leaves. Glycoconjugate J 13, 977–983 (1996). https://doi.org/10.1007/BF01053193

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01053193

Keywords

Navigation