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

Journal Article · Tue Jan 14 00:00:00 EST 1992 · Biochemistry; (United States)

DOI:https://doi.org/10.1021/bi00116a039· OSTI ID:5487492

Ip, C C.Y.; Miller, W J; Kubek, D J ^[1]; Strang, A M; van Halbeek, H ^[2]; Piesecki, S J; Alhadeff, J A ^[3]

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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)

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.

• OSTI ID: 5487492
• Journal Information: Biochemistry; (United States), Vol. 31:1; ISSN 0006-2960
• Country of Publication: United States
• Language: English

Related Subjects

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