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N-acetylglucosaminyltransferase substrates prepared from glycoproteins by hydrazinolysis of the asparagine-N-acetylglucosamine linkage. Purification and structural determination of oligosaccharides with mannose andN-acetylglucosamine at the non-reducing termini

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Abstract

Sixteen asparagine-linked oligosaccharides ranging in size from (Man)2(GlcNAc)2 (Fuc)1 to (GlcNAc)6(Man)3(GlcNAc)2 were obtained from human α1-acid glycoprotein and fibrinogen, hen ovomucoid and ovalbumin, and bovine fetuin, fibrin and thyroglobulin by hydrazinolysis, mild acid hydrolysis and glycosidase treatment. The oligosaccharides hadN-acetylglucosamine at the reducing termini and mannose andN-acetylglucosamine residues at the non-reducing termini and were prepared for use asN-acetylglucosaminyltransferase substrates. Purification of the oligosaccharides involved gel filtration and high performance liquid chromatography on reverse phase and amine-bonded silica columns. Structures were determined by 360 MHz and 500 MHz proton nuclear magnetic resonance spectroscopy, fast atom bombardment-mass spectrometry and methylation analysis. Several of these oligosaccharides have not previously been well characterized.

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Abbreviations

bis:

bisecting GlcNAc

DMSO:

dimethylsulfoxide

FAB:

fast atom bombardment

Fuc:

l-fucose

Gal:

d-galactose

GLC:

gas-liquid chromatography

GlcNAc or Gn:

N-acetyl-d-glucosamine

HPLC:

high performance liquid chromatography

Man or M:

d-mannose

MES:

2-(N-morpholino)ethanesulfonate

MS:

mass spectrometry

NMR:

nuclear magnetic resonance

PIPES:

piperazine-N,N′-bis(2-ethane sulfonic acid)

References

  1. Yamashita K, Ohkura T, Tachibana Y, Takasaki S, Kobata A (1984) J Biol Chem 259:10834–40.

    Google Scholar 

  2. Pierce M, Arango J (1986) J Biol Chem 261:10772–77.

    Google Scholar 

  3. Dennis JW, Laferte S, Waghorne C, Breitman ML, Kerbel RS (1987) Science 236:582–85.

    Google Scholar 

  4. Narasimhan S, Schachter H, Rajalakshmi S (1988) J Biol Chem 263:1273–81.

    Google Scholar 

  5. Schachter H (1986) Biochem Cell Biol 64:163–81.

    Google Scholar 

  6. Brockhausen I, Carver JP, Schachter H (1988) Biochem Cell Biol, in press.

  7. Brockhausen I, Grey AA, Carver JP, Hindsgaul O, Schachter H (1987) Proc IXth Int Symp Glycoconjugates, eds. Montreuil J, Verbert A, Spik G, Fournet B, Secretariat, Lille, E35.

    Google Scholar 

  8. Li Y-T, Li S-C (1972) Methods Enzymol 28:702–13.

    Google Scholar 

  9. Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F (1956) Anal Chem 28: 350–56.

    Google Scholar 

  10. Svennerholm L (1958) Acta Chem Scand 12:547–54.

    Google Scholar 

  11. Zanetta JP, Breckenridge WC, Vincendon G (1972) J Chromatogr 69:291–304.

    Google Scholar 

  12. Bendiak B, Cumming DA (1985) Carbohydr Res 144:1–12.

    Google Scholar 

  13. Bendiak B, Cumming DA (1986) Carbohydr Res 151:89–103.

    Google Scholar 

  14. Narasimhan S, Harpaz N, Longmore G, Carver JF, Grey AA, Schachter H (1980) J Biol Chem 255:4876–84.

    Google Scholar 

  15. Narasimhan S, Freed JC, Schachter H (1985) Biochemistry 24:1694–1700.

    Google Scholar 

  16. Distler JJ, Jourdian GW (1978) Methods Enzymol 50:514–20.

    Google Scholar 

  17. Plummer TH, Tarentino AL (1981) J Biol Chem 256:10243–46.

    Google Scholar 

  18. Ciucanu I, Kerek F (1984) Carbohydr Res 131:209–17.

    Google Scholar 

  19. Waeghe TJ, Darvill AG, McNeil M, Albersheim P (1983) Carbohydr Res 123:281–304.

    Google Scholar 

  20. Carver JP, Grey AA (1981) Biochemistry 20:6607–16.

    Google Scholar 

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

    Google Scholar 

  22. Grey AA, Narasimhan S, Brisson J-R, Schachter H, Carver JP (1982) Can J Biochem 60:1123–31.

    Google Scholar 

  23. Gleeson PA, Schachter H (1983) J Biol Chem 258:6162–73.

    Google Scholar 

  24. Schut BL, Dorland L, Haverkamp J, Vliegenthart JFG, Fournet B (1978) Biochem Biophys Res Comm 82:1223–28.

    Google Scholar 

  25. Dorland L, Haverkamp J, Vliegenthart JFG, Spik G, Fournet B, Montreuil J (1979) Eur J Biochem 100:569–74.

    Google Scholar 

  26. Ceccarini C, Lorenzoni P, Atkinson P (1983) Biochim Biophys Acta 759:214–21.

    Google Scholar 

  27. Paz-Parente J, Wieruszeski J-M, Strecker G, Montreuil J, Fournet B van Halbeek H, Dorland L, Vliegenthart JFG (1982) J Biol Chem 257:13173–76.

    Google Scholar 

  28. Carver JP, Grey AA, Winnik FM, Hakimi J, Ceccarini C, Atkinson P (1981) Biochemistry 20:6600–6.

    Google Scholar 

  29. Townsend RR, Hilliker E, Li Y-T, Laine R, Bell WR, Lee YC (1982) J Biol Chem 257:9704–10.

    Google Scholar 

  30. Ito S, Yamashita K, Spiro RG, Kobata A (1977) J Biochem (Tokyo) 81:1621–31.

    Google Scholar 

  31. Nilsson B, Nordén NE, Svensson S (1979) J Biol Chem 254:4545–53.

    Google Scholar 

  32. Baenzinger JU, Fiete D (1979) J Biol Chem 254:789–95.

    Google Scholar 

  33. Fournet B, Montreuil J, Strecker G, Dorland L, Haverkamp J, Vliegenthart JFG, Binette JP, Schmid K (1978) Biochemistry 17:5206–14.

    Google Scholar 

  34. van Halbeek H, Dorland L, Veldink GA, Vliegenthart JFG, Michalski J-C, Montreuil J, Strecker G, Hull WE (1980) FEBS Lett 121:65–70.

    Google Scholar 

  35. Yamashita K, Kamerling JP, Kobata A (1982) J Biol Chem 257:12809–14.

    Google Scholar 

  36. Yamashita K, Kamerling JP, Kobata A (1983) J Biol Chem 258:3099–106.

    Google Scholar 

  37. Paz-Parente J, Strecker G, Leroy Y, Montreuil J, Fournet B, van Halbeek H, Dorland L, Vliegenthart JFG (1983) FEBS Lett 152:145–52.

    Google Scholar 

  38. Egge H, Peter-Katalinic J, Paz-Parente J, Strecker G, Montreuil J, Fournet B (1983) FEBS Lett 156:357–62.

    Google Scholar 

  39. Geyer R, Diabate S, Geyer H, Klenk H-D, Niemann H, Stirm S (1987) Glycoconjugate J 4:17–32.

    Google Scholar 

  40. van Halbeek H, Vliegenthart JFG, Iwase H, Li S-C, Li Y-T (1985) Glycoconjugate J 2:235–53.

    Google Scholar 

  41. Stanley P (1985) Mol Cell Biol 5:923–29.

    Google Scholar 

  42. Deutscher SL, Hirschberg CB (1986) J Biol Chem 261:96–100.

    Google Scholar 

  43. Dennis JW, Carver JP, Schachter H (1984) J Cell Biol 99:1034–44.

    Google Scholar 

  44. Dennis JW, Laferte S, Fukuda M, Dell A, Carver JP (1986) Eur J Biochem 161:359–73.

    Google Scholar 

  45. Narasimhan S (1982) J Biol Chem 257:10235–42.

    Google Scholar 

  46. Cummings RD, Trowbridge IS, Kornfeld S (1982) J Biol Chem 257:13421–27.

    Google Scholar 

  47. Koenderman AHL, Wijermans PW, van den Eijnden DH (1987) FEBS Lett 222:42–46.

    Google Scholar 

  48. Oppenheimer CL, Hill RL (1981) J Biol Chem 256:799–804.

    Google Scholar 

  49. Vella GJ, Paulsen H, Schachter H (1984) Can J Biochem Cell Biol 62:409–17.

    Google Scholar 

  50. Bendiak B, Schachter H (1987) J Biol Chem 262:5784–90.

    Google Scholar 

  51. Sturm A, van Kuik JA, Vliegenthart JFG, Chrispeels MJ (1987) J Biol Chem 262:13392–403.

    Google Scholar 

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Authors and Affiliations

  1. Research Institute, Hospital for Sick Children, 555 University Avenue, M5G 1X8, Toronto, Ontario, Canada

    Inka Brockhausen & Harry Schachter

  2. Department of Medical Genetics and Medical Biophysics, University of Toronto, M5S 1A8, Toronto, Ontario, Canada

    Inka Brockhausen, Arthur A Grey, Henrianna Pang & Jeremy P Carver

  3. Department of Biochemistry, University of Toronto, M5S 1A8, Toronto, Ontario, Canada

    Harry Schachter

Authors
  1. Inka Brockhausen
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  2. Arthur A Grey
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  3. Henrianna Pang
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  4. Harry Schachter
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  5. Jeremy P Carver
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Additional information

the nomenclature of the oligosaccharides is shown in Table 1.

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Brockhausen, I., Grey, A.A., Pang, H. et al. N-acetylglucosaminyltransferase substrates prepared from glycoproteins by hydrazinolysis of the asparagine-N-acetylglucosamine linkage. Purification and structural determination of oligosaccharides with mannose andN-acetylglucosamine at the non-reducing termini. Glycoconjugate J 5, 419–448 (1988). https://doi.org/10.1007/BF01049917

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  • DOI: https://doi.org/10.1007/BF01049917

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