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Title: Structural characterization of the N-glycans of a recombinant hepatitis B surface antigen derived from yeast

Abstract

The N-glycans of purified recombinant middle surface protein (preS2+S) from hepatitis B virus, a candidate vaccine antigen expressed in a mnn9 mutant strain of Saccharomyces cerevisiae, have been characterized structurally. The glycans were released by N-glycanase treatment, isolated by size-exclusion chromatography on Sephadex G-50 and Bio-Gel P-4 columns, and analyzed by 500-MHz {sup 1}H NMR spectroscopy and fast atom bombardment mass spectrometry. The mixture of oligosaccharides was fractionated by HPLC, the major subfractions were isolated, and their carbohydrate compositions were determined by high-pH anion-exchange chromatography with pulsed amperometric detection. The combined results suggest that high-mannose oligosaccharides account for all the N-glycans released from preS2+S: structures include Man{sub 7}GlcNAc{sub 2}, Man{sub 8}GlcNAc{sub 2} isomers in the ratios of 3:6:1. Approximately 80% of the oligosaccharides contain the C2, C6-branched trimannosyl structural element typical of yeast high-mannose oligosaccharides but not usually found in high-mannose oligosaccharides in animal glycoproteins.

Authors:
; ;  [1]; ;  [2]; ;  [3]
  1. Merck Sharp and Dohme Research Labs., West Point, PA (United States)
  2. Univ. of Georgia, Athens (United States)
  3. Lehigh Univ., Bethlehem, PA (United States)
Publication Date:
OSTI Identifier:
5487492
Resource Type:
Journal Article
Journal Name:
Biochemistry; (United States)
Additional Journal Information:
Journal Volume: 31:1; Journal ID: ISSN 0006-2960
Country of Publication:
United States
Language:
English
Subject:
62 RADIOLOGY AND NUCLEAR MEDICINE; ANTIGENS; NUCLEAR MAGNETIC RESONANCE; HEPATITIS; IMMUNOLOGY; MEMBRANE PROTEINS; MOLECULAR STRUCTURE; GLYCOPROTEINS; MASS SPECTROSCOPY; SACCHAROMYCES CEREVISIAE; VACCINES; DIGESTIVE SYSTEM DISEASES; DISEASES; EUMYCOTA; FUNGI; MAGNETIC RESONANCE; MICROORGANISMS; ORGANIC COMPOUNDS; PLANTS; PROTEINS; RESONANCE; SACCHAROMYCES; SPECTROSCOPY; YEASTS; 550601* - Medicine- Unsealed Radionuclides in Diagnostics

Citation Formats

Ip, C C.Y., Miller, W J, Kubek, D J, Strang, A M, van Halbeek, H, Piesecki, S J, and Alhadeff, J A. Structural characterization of the N-glycans of a recombinant hepatitis B surface antigen derived from yeast. United States: N. p., 1992. Web. doi:10.1021/bi00116a039.
Ip, C C.Y., Miller, W J, Kubek, D J, Strang, A M, van Halbeek, H, Piesecki, S J, & Alhadeff, J A. Structural characterization of the N-glycans of a recombinant hepatitis B surface antigen derived from yeast. United States. doi:10.1021/bi00116a039.
Ip, C C.Y., Miller, W J, Kubek, D J, Strang, A M, van Halbeek, H, Piesecki, S J, and Alhadeff, J A. Tue . "Structural characterization of the N-glycans of a recombinant hepatitis B surface antigen derived from yeast". United States. doi:10.1021/bi00116a039.
@article{osti_5487492,
title = {Structural characterization of the N-glycans of a recombinant hepatitis B surface antigen derived from yeast},
author = {Ip, C C.Y. and Miller, W J and Kubek, D J and Strang, A M and van Halbeek, H and Piesecki, S J and Alhadeff, J A},
abstractNote = {The N-glycans of purified recombinant middle surface protein (preS2+S) from hepatitis B virus, a candidate vaccine antigen expressed in a mnn9 mutant strain of Saccharomyces cerevisiae, have been characterized structurally. The glycans were released by N-glycanase treatment, isolated by size-exclusion chromatography on Sephadex G-50 and Bio-Gel P-4 columns, and analyzed by 500-MHz {sup 1}H NMR spectroscopy and fast atom bombardment mass spectrometry. The mixture of oligosaccharides was fractionated by HPLC, the major subfractions were isolated, and their carbohydrate compositions were determined by high-pH anion-exchange chromatography with pulsed amperometric detection. The combined results suggest that high-mannose oligosaccharides account for all the N-glycans released from preS2+S: structures include Man{sub 7}GlcNAc{sub 2}, Man{sub 8}GlcNAc{sub 2} isomers in the ratios of 3:6:1. Approximately 80% of the oligosaccharides contain the C2, C6-branched trimannosyl structural element typical of yeast high-mannose oligosaccharides but not usually found in high-mannose oligosaccharides in animal glycoproteins.},
doi = {10.1021/bi00116a039},
journal = {Biochemistry; (United States)},
issn = {0006-2960},
number = ,
volume = 31:1,
place = {United States},
year = {1992},
month = {1}
}