1887

Journal of General Virology header logo

Volume 71, Issue 4

Differential glycosylation of the protein (PrP) forming scrapie-associated fibrils Free

Abstract

PrP is a glycoprotein found in normal brain. In brain affected by scrapie it forms scrapie-associated fibrils (SAF). PrP from SAF shows considerable heterogeneity of size and charge on two-dimensional gels. It separates into six major regions, the three more acidic regions arising as a result of partial proteolytic degradation. The two more basic higher forms ( 34000 and 29000) of PrP can be reduced in apparent to a lower form ( 25000) with Peptide-- glycosidase F. In addition, a series of lectins has been found to bind to PrP. Some bind preferentially to the higher forms whereas others bind more strongly to the lower form. Some of the heterogeneity of PrP is therefore due to differential -glycosylation. We suggest that one or two -linked carbohydrate chains are bound to the protein causing some of the differences in The major cause of heterogeneity of PrP is therefore proteolytic cleavage combined with differential glycosylation at the two potential -glycosylation sites. The glycolipid moiety attached to PrP may be responsible for some lectin binding to all three bands. Using lectins as a probe to study potential differences in -glycosylation we have looked at their binding to PrP isolated from SAF, from different strains of scrapie and from different regions of the same brain. No major differences in the -glycan moieties were found.

  • Received:
  • Accepted:
  • Published Online:
© Society for General Microbiology 1990
Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-71-4-833
1990-04-01
2024-04-27
Loading full text...

Full text loading...

/deliver/fulltext/jgv/71/4/JV0710040833.html?itemId=/content/journal/jgv/10.1099/0022-1317-71-4-833&mimeType=html&fmt=ahah

References

  1. Bause E. 1979; Studies on the acceptor specificity of asparagine-N-glycosyl-transferase from rat liver. FEBS Letters 103:296–300
     [Google Scholar]
  2. Bolton D. C., Mckinley M. P., Prusiner S. B. 1982; Identification of a protein that purifies with the scrapie prion. Science 218:1309–1311
     [Google Scholar]
  3. Bolton D. C., Meyer R. K., Prusiner S. B. 1985; Scrapie PrP 27-30 is a sialoglycoprotein. Journal of Virology 53:596–606
     [Google Scholar]
  4. Caughey B., Race R. E., Ernst D., Buchmeier M. J., Chesebro B. 1989; Prion protein biosynthesis in scrapie-infected and uninfected neuroblastoma cells. Journal of Virology 63:175–181
     [Google Scholar]
  5. Dickinson A. G., Meikle V. M. H., Fraser H. 1968; Identification of a gene which controls the incubation period of some strains of scrapie agent in mice. Journal of Comparative Pathology 78:293–299
     [Google Scholar]
  6. Diringer H., Gelderblom H., Hilmert H., Ozel M., Edelbluth C., Kimberlin R. H. 1983; Scrapie infectivity, fibrils and low molecular weight protein. Nature; London: 306476–478
     [Google Scholar]
  7. Haraguchi T., Fisher S., Olofsson S., Endo T., Groth D., Tarentino A., Borchelt D. R., Teplow D., Hood L., Burlingame A., Lycke E., Kobata A., Prusiner S. B. 1989; Asparagine-linked glycosylation of the scrapie and cellular prion proteins. Archives of Biochemistry and Biophysics 274:1–13
     [Google Scholar]
  8. Hope J., Morton L. J. D., Farquhar C. F., Multhaup G., Beyreuther K., Kimberlin R. H. 1986; The major polypeptide of scrapie-associated fibrils (SAF) has the same size, charge distribution and N-terminal protein sequence as predicted for the normal brain protein (PrP). EMBO Journal 5:2591–2597
     [Google Scholar]
  9. Hope J., Multhaup G., Reekie L. J. D., Kimberlin R. H., Beyreuther K. 1988; Molecular pathology of scrapie-associated fibril protein (PrP) in mouse brain affected by the ME7 strain of scrapie. European Journal of Biochemistry 172:271–277
     [Google Scholar]
  10. Hunkapillar M. W., Lujan E., Ostrander F., Hood L. E. 1983; Electroelution methods in enzymology. Methods in Enzymology 91:221–236
     [Google Scholar]
  11. Hunter N., Hope J., Mcconnell I., Dickinson A. G. 1987; Linkage of the scrapie-associated fibril protein (PrP) gene and Sinc using congenic mice and restriction fragment length polymorphism analysis. Journal of General Virology 68:2711–2716
     [Google Scholar]
  12. Kascsak R. J., Rubenstein R., Merz P. A., Carp R. I., Robakis N. K., Wisniewski H. M., Diringer H. 1986; Immunological comparison of scrapie-associated fibrils isolated from animals infected with four different strains of scrapie. Journal of Virology 59:676–683
     [Google Scholar]
  13. Manuelidis L., Valley S., Manuelidis E. E. 1985; Specific proteins associated with Creutzfeldt-Jakob disease and scrapie share antigenic and carbohydrate determinants. Proceedings of the National Academy of Sciences, U.S.A 82:4263–4267
     [Google Scholar]
  14. Merz P. A., Somerville R. A., Wisniewski H. M., Iqbal I. 1981; Abnormal fibrils from scrapie-infected brain. Acta neuropathologica 54:63–74
     [Google Scholar]
  15. Merz P. A., Somerville R. A., Wisniewski H. M., Manuelidis L., Manuelidis E. E. 1983; Scrapie-associated fibrils in Creutzfeld-Jakob disease. Nature; London: 306474–476
     [Google Scholar]
  16. Merz P. A., Rohwer R. G., Kascsak R., Wisniewski H. M., Somerville R. A., Gibbs C. J., Gajdusek D. C. 1984; An infection-specific particle from the unconventional virus diseases. Science 225:437–440
     [Google Scholar]
  17. Merz P. A., Kascsak R. J., Rubenstein R., Carp R. I., Wisniewski H. M. 1987; Antisera to scrapie-associated fibril protein and prion protein decorate scrapie-associated fibrils. Journal of Virology 61:42–49
     [Google Scholar]
  18. Multhaup G., Diringer H., Hilmert H., Prinz H., Heukeshoven J., Beyreuther K. 1985; The protein component of scrapie- associated fibrils is a glycosylated low molecular weight protein. EMBO Journal 4:1495–1501
     [Google Scholar]
  19. Neville D. M. 1971; Molecular weight determination of protein-dodecyl sulphate complexes by gel electrophoresis in a discontinuous buffer system. Journal of Biological Chemistry 246:6328–6334
     [Google Scholar]
  20. Oesch B., Westaway D., Walchi M., Mckinley M. P., Kent S. B. H., Aebersold R., Barry R. A., Tempst P., Teplow D. B., Hood L. E., Prusiner S. B., Weissmann C. 1985; A cellular gene encodes scrapie PrP27-30 protein. Cell 40:735–746
     [Google Scholar]
  21. O’Farrell P. Z., Goodman H. M., O’Farrell P. H. 1977; High resolution two-dimensional electrophoresis of basic as well as acidic proteins. Cell 12:1133–1142
     [Google Scholar]
  22. Plummer T. H., Elder J. H., Alexander S., Phelan A. W., Tarentino A. L. 1984; Demonstration of peptide: N-glycosidase F activity in endo-β-N-acetylglucosaminidase F preparations. Journal of Biological Chemistry 259:10700–10704
     [Google Scholar]
  23. Sammons D. W., Adams L. D., Nishizawa E. E. 1981; Ultrasensitive silver-based color staining of polypeptides in polyacrylamide gels. Electrophoresis 2:135–141
     [Google Scholar]
  24. Sharon N., Lis H. 1989 Lectins London: Chapman and Hall;
     [Google Scholar]
  25. Sklaviadis T., Manuelidis L., Manuelidis E. E. 1986; Characterisation of major peptides in Creutzfeldt-Jakob disease and scrapie. Proceedings of the National Academy of Sciences, U.S.A 83:6146–6150
     [Google Scholar]
  26. Somerville R. A., Ritchie L. A. 1989; Are scrapie associated fibrils a pathological product of infection?. In Unconventional Virus Diseases of the Central Nervous System pp. 521–535 Court L. A., Dormont D., Brown P., Kingsbury D. T. Edited by Fontenay-aux-Roses: Commissariat à l’Energie Atomique;
     [Google Scholar]
  27. Somerville R. A., Merz P. A., Carp R. I. 1986; Partial copurification of scrapie-associated fibrils and scrapie infectivity. Intervirology 25:48–55
     [Google Scholar]
  28. Somerville R. A., Ritchie L. A., Gibson P. H. 1989; Structural and biochemical evidence that scrapie-associated fibrils assemble in vivo. Journal of General Virology 70:25–35
     [Google Scholar]
  29. Stahl N., Borchelt D. R., Hsiao K., Prusiner S. B. 1987; Scrapie prion protein contains a phosphatidyl-inositol glycolipid. Cell 51:229–240
     [Google Scholar]
  30. Towbin H., Staehelin T., Gordon J. 1979; Electrophoretic transfer of protein from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences, U.S.A 76:4350–4354
     [Google Scholar]
  31. Westaway D., Goodman P. A., Mirenda C. A., Mckinley M. P., Carlson G. A., Prusiner S. B. 1987; Distinct prion proteins in short and long incubation period mice. Cell 51:651–662
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-71-4-833
Loading
Differential glycosylation of the protein (PrP) forming scrapie-associated fibrils
J. Gen. Virol. 71, 833 (1990); https://doi.org/10.1099/0022-1317-71-4-833
/content/journal/jgv/10.1099/0022-1317-71-4-833
/content/journal/jgv/10.1099/0022-1317-71-4-833
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error