Abstract
The effects of treatments of the glycoprotein ribonuclease-B, the proteins ribonuclease-A and myoglobin, and the glyco-amino acid GlcNAcβ(1-N) Asn with alkalil alkaline sodium borohydride, and aqueous sodium borohydride were systematically studied as a function of the concentration of the reagents, the temperature, and the length of the treatment. High-field1H-NMR spectroscopy, chromatographic methods and amino-acid analysis were used to characterize products of the treatments of the various compounds. Our results indicate that mild alkaline borohydride treatment, as well as aqueous borohydride treatment alone, is capable of extensively degrading polypeptides and of partially releasing theN-linked glycans from ribonuclease-B. Initially, glycopeptides are produced, the peptide portion of which consists of several amino acids, which are further hydrolyzed to yield a mixture of glyco-asparagines and oligosaccharide-alditols in the ratio of ≈4:1. Strong alkaline borohydride treatment of ribonuclease-B is capable of completely releasing theN-linked carbohydrates as oligosaccharide-alditols.
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Abbreviations
- RNase:
-
ribonuclease
References
Spiro RG (1972) Methods Enzymol 28:32–34
Sharon N, Lis H (1982) in Proteins, vol 5, eds. Neurath H, Hill R, Academic Press, New York, p 1–144.
Zinn AB, Marshall JS, Carlson DM (1978) J Biol Chem 253:6761–67.
Rasilo M-J, Renkonen O (1981) FEBS Lett 135:38–42.
Ogata S-I, Lloyd KO (1982) Anal Biochem 119:351–59.
Debray H, Strecker G, Montreuil J (1984) Biochem Soc Trans 12:611–12.
lyer RN, Carlson DM (1971) Arch Biochem Biophys 142:101–5.
Spiro RG, Bhoyroo VD (1974) J Biol Chem 249:5704–17.
Hounsell EF, Pickering NJ, Stoll MS, Lawson AM, Feizi T (1984) Biochem Soc Trans 12:607–10.
Bernard BA, Newton SA, Olden K (1983) J Biol Chem 258:12198–202.
Hewitt LF (1937) Biochem J 31:360–66.
Vliegenthart JFG, Dorland L, van Halbeek H (1983) Adv Carbohydr Chem Biochem 41:209–374.
Sinha S, Watorek W, Karr S, Giles J, Bode W, Travis J (1987) Proc Natl Acad Sci USA 84:2228–32.
Ronin C, van Halbeek H, Mutsaers JHGM, Vliegenthart JFG (1987) Glycoconjugate J 4:247–54.
Mutsaers JHGM, van Halbeek H, Vliegenthart JFG, Tager JM, Reuser AJJ, Kroos M, Galjaard H (1987) Biochim Biophys Acta 911:244–51.
Liang C-J, Yamashita K, Kobata A (1980) J Biochem (Tokyo) 88:51–58.
Marti T, Kosen PA, Honory K, Franzblau C, Schmid K, van Halbeek H, Gerwig GJ, Vliegenthart JFG (1984) Biochim Biophys Acta 799:305–12.
Mutsaers JHGM, van Halbeek H, Kamerling JP, Vliegenthart JFG (1985) Eur J Biochem 147:569–74.
Overdijk B, Beem EP, van Steijn GJ, Trippelvitz LAW, Lisman JJW, Paz-Parente J, Cardon P, Leroy Y, Fournet B, van Halbeek H, Mutsaers JHGM, Vliegenthart JFG (1985) Biochem J 232:637–41.
Ogata S, Misumi Y, Miki K, Ikehara Y (1986) Eur J Biochem 161:315–20.
Takasaki S, Mizuochi T, Kobata A (1982) Methods Enzymol 83:263–68.
Michalski JC, Peter-Katalinic J, Egge H, Paz-Parente J, Montreuil J, Strecker G (1984) Carbohydr Res 134:177–89.
Lloyd KO, Kabat EA, Licerio E (1968) Biochemistry 7:2976–90.
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Argade, S.P., Daves, G.D., Van Halbeek, H. et al. The effect of alkaline borohydride treatment onN-linked carbohydrates of glycoproteins. Glycoconjugate J 6, 45–56 (1989). https://doi.org/10.1007/BF01047889
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DOI: https://doi.org/10.1007/BF01047889