Advertisement

Glycoconjugate Journal

, Volume 10, Issue 1, pp 64–71 | Cite as

Human erythrocyte sialidase is linked to the plasma membrane by a glycosylphosphatidylinositol anchor and partly located on the outer surface

  • Anna Chiarini
  • Amelia Fiorilli
  • Lucia Di Francesco
  • Bruno Venerando
  • Guido Tettamanti
Papers

Abstract

Treatment of human erythrocyte ghosts with phosphatidylinositol-phospholipase C (PIPLC) fromBacillus cereus liberated the ghost-linked sialidase. Maximal release of sialidase (about 70% of total) was achieved by incubating ghosts at 37°C for 60 min, at pH 6.0, with PIPLC (PIPLC total units/ghost protein ratio, 4.5 each time) added at the beginning of incubation and every 15 min (four subsequent additions). Liberated sialidase was fully resistant to at least four cycles of rapid freezing and thawing and to storage at 4°C for at least 48 h. The liberated enzyme had an optimal activity at pH 4.2, degraded ganglioside GD1a better than methylumbelliferylN-acetylneuraminic acid (about fourfold), and gave aKm value of 2.56 · 10−4m and an apparentVmax of 2.22 mU per mg protein on GD1a. Treatment of intact erythrocytes with PIPLC (PIPLC total units/erythrocyte protein ratio, 8), under conditions where haemolysis was practically negligible, caused liberation of 10–12% of membrane linked sialidase, indicating that the enzyme is, at least in part, located on the outer surface of the erythrocyte membrane. It is concluded that the erythrocyte membrane sialidase is anchored by a glycosylphosphatidylinositol structure sensitive to PIPLC action, and is partly located on the outer surface.

Keywords

erythrocyte sialidase glycosylphosphatidylinositol anchor exoenzymes 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Aminoff D, Bell WC, Fulton I, Ingebrigtsen N (1976)Am J Hematol 1: 419–32.Google Scholar
  2. 2.
    Bell WC, Levy GN, Williams K, Aminoff D (1977)Proc Natl Acad Sci USA 74: 4205–9.Google Scholar
  3. 3.
    Kazuyuki T, Makoto M (1988)Toxicology 53: 269–76.Google Scholar
  4. 4.
    Kuester JM, Schauer R (1981)Hoppe Seyl Z Physiol Chem 362: 1507–14.Google Scholar
  5. 5.
    Mueller E, Schroeder C, Schauer R, Sharon N (1983)Hoppe Seyl Z Physiol Chem 364: 1419–29.Google Scholar
  6. 6.
    Franco M, Nordt FJ, Corfield AP, Schauer R (1981)Acta Biol Med Gen 40: 409–12.Google Scholar
  7. 7.
    Gutowski KA, Linseman DA, Aminoff D (1988)Carbohydr Res 178: 307–13.Google Scholar
  8. 8.
    Cabezas JA (1991)Biochem J 278: 311–2.Google Scholar
  9. 9.
    Schengrund CL, Rosenberg A (1970)J Biol Chem 245: 6196–200.Google Scholar
  10. 10.
    Tettamanti G, Morgan IG, Gombos G, Vincendon G, Mandel P (1972)Brain Res 47: 575–8.Google Scholar
  11. 11.
    Venerando B, Tettamanti G, Cestaro B, Zambotti V (1975)Biochim Biophys Acta 403: 461–72.Google Scholar
  12. 12.
    Miyagi T, Tsuiki S (1985)J Biol Chem 260: 6710–6.Google Scholar
  13. 13.
    Fiorilli A, Venerando B, Siniscalco C, Monti E, Bresciani R, Caimi L, Preti A, Tettamanti G (1989)J Neurochem 53: 672–80.Google Scholar
  14. 14.
    Gielen W, Etzrodt J, Uhlenbruck G (1969)Thromb Diath Haemorrh 22: 203–7.Google Scholar
  15. 15.
    Bosmann HB (1974)Vox Sang 26: 497–512.Google Scholar
  16. 16.
    Sagawa J, Miyagi T, Tsuiki S (1990)J Biochem (Tokyo)107: 452–6.Google Scholar
  17. 17.
    Slein MW, Logan GK (1985)J Bacteriol 90: 69–81.Google Scholar
  18. 18.
    Tettamanti G, Bonali F, Marchesini S, Zambotti V (1973)Biochim Biophys Acta 296: 160–70.Google Scholar
  19. 19.
    Sonnino S, Ghidoni R, Galli G, Tettamanti G (1978)J Neurochem 31:947–56.Google Scholar
  20. 20.
    Ghidoni R, Sonnino S, Masserini M, Orlando P, Tettamanti G (1981)J Lipid Res 22: 1286–95.Google Scholar
  21. 21.
    Sonnino S, Ghidoni R, Gazzotti G, Kirschner G, Galli G, Tettamanti G (1984)J Lipid Res 25: 620–9.Google Scholar
  22. 22.
    Ikezawa M, Yamanagi M, Taguchi R, Miyashita T, Ohyabu T (1976)Biochim Biophys Acta 450: 154–64.Google Scholar
  23. 23.
    Low MG, Finean JB (1977)FEBS Lett 82: 143–6.Google Scholar
  24. 24.
    Taguchi R, Suzuki K, Nakabayashi T, Ikezawa H (1984)J Biochem (Tokyo)96: 437–46.Google Scholar
  25. 25.
    Taguchi R, Mizuno M, Inoue M, Ikezawa H (1983)J Biochem (Tokyo)93: 403–12.Google Scholar
  26. 26.
    Futerman AH, Low MG, Michaelson PM, Silman I (1985)J Neurochem 45: 1487–94.Google Scholar
  27. 27.
    Marchesini S, Venerando B, Fiorilli A, Preti A (1981)Perspect Inherited Metab Dis 4: 191–203.Google Scholar
  28. 28.
    Chigorno V, Cardace G, Pitto M, Sonnino S, Ghidoni R, Tettamanti G (1986)Anal Biochem 183: 283–94.Google Scholar
  29. 29.
    Ellman GL, Courtney DK, Andres V, Featherstone RM (1961)Biochem Pharmacol 7: 88–95.Google Scholar
  30. 30.
    Ghidoni R, Trinchera M, Venerando B, Fiorilli A, Sonnino S, Tettamanti G (1986)Biochem J 237: 147–55.Google Scholar
  31. 31.
    Sonnino S, Kirschner G, Fronza G, Egge H, Ghidoni R, Acquotti D, Tettamanti G (1985)Glycoconjugate J 2: 343–54.Google Scholar
  32. 32.
    Lowry OM, Rosebrough NJ, Farr AL, Randall RJ (1951)J Biol Chem 193: 265–75.Google Scholar
  33. 33.
    Bradford MM (1976)Anal Biochem 72: 248–54.Google Scholar
  34. 34.
    Svennerholm L (1957)Biochim Biophys Acta 24: 604–11.Google Scholar
  35. 35.
    Low MG, Finean JB (1977)Biochem J 162: 235–40.Google Scholar
  36. 36.
    Butikofer P, Lin ZW, Chiu DTY, Lubin B, Kuypers FA (1990)J Biol Chem 265: 16035–8.Google Scholar
  37. 37.
    Low MG, Finean JB (1978)Biochim Biophys Acta 508: 565–70.Google Scholar
  38. 38.
    Ferguson AJ (1988)Annu Rev Biochem 57: 285–320.Google Scholar
  39. 39.
    Chiarini A, Fiorilli A, Siniscalco C, Tettamanti G, Venerando B (1990)J Neurochem 55: 1576–84.Google Scholar
  40. 40.
    Carrol PT, Smith LK (1990)J Neurochem 54: 1047–55.Google Scholar
  41. 41.
    Shukla SD, Coleman R, Finean JB, Michell RH (1980)Biochem J 187: 277–80.Google Scholar
  42. 42.
    Svennerholm L (1980)Adv Exptl Med Biol 125: 11–22.Google Scholar
  43. 43.
    IUPAC-IUB Commission on Biochemical Nomenclature (1977)Lipids 12:455–68; (1982)J Biol Chem 257:3347–51.Google Scholar

Copyright information

© Chapman & Hall 1993

Authors and Affiliations

  • Anna Chiarini
    • 1
  • Amelia Fiorilli
    • 1
  • Lucia Di Francesco
    • 1
  • Bruno Venerando
    • 1
  • Guido Tettamanti
    • 1
  1. 1.Department of Medical Chemistry and Biochemistry, The Medical SchoolUniversity of MilanMilanItaly

Personalised recommendations