Document Detail

Primary structure of 21 novel monoantennary and diantennary N-linked carbohydrate chains from alphaD-hemocyanin of Helix pomatia.
MedLine Citation:
PMID:  9363772     Owner:  NLM     Status:  MEDLINE    
The primary structures of 21 novel monoantennary and diantennary N-glycans of the glycoprotein alphaD-hemocyanin (alphaD-Hc) of Helix pomatia have been determined. Outer oligosaccharide fragments (antennae) were released from the glycoprotein by Smith degradation of an alphaD-Hc pronase digest. The major antenna, obtained following HPLC fractionation on Lichrosorb-NH2, was characterized using 1H-NMR spectroscopy, fast-atom-bombardment mass spectrometry, and linkage analysis, and corresponds to a pentasaccharide fragment. The intact carbohydrate chains of alphaD-Hc were released with peptide-N4-(N-acetyl-beta-glucosaminyl)asparagine amidase-F digestion, separated from the protein on Bio-Gel P-100, and subfractionated on Bio-Gel P-4. A portion of subfractions was reduced with sodium borodeuteride, and the non-reduced and reduced samples were further fractionated on CarboPac PA-1, Lichrosorb-NH2/Lichrosphere-NH2, and/or Lichrosphere-C18. Purified oligosaccharides and oligosaccharide-alditols were analyzed using 500/600-MHz 1H-NMR spectroscopy. In total, four novel types of antenna were identified, namely, [structures: see text] which are all attached to O-2 of alphaMan residues of the trimannosyl-N,N'-diacetylchitobiose core element, which is generally beta-1,2-xylosylated and alpha-1,6-fucosylated, Man(alpha1-6)[Man(alpha1-3)][+/-Xyl(beta1-2)]Man(beta1-4)GlcNAc(beta1-4) [+/-Fuc(alpha1-6)]GlcNAc.
J P Lommerse; J E Thomas-Oates; C Gielens; G Préaux; J P Kamerling; J F Vliegenthart
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  European journal of biochemistry / FEBS     Volume:  249     ISSN:  0014-2956     ISO Abbreviation:  Eur. J. Biochem.     Publication Date:  1997 Oct 
Date Detail:
Created Date:  1997-12-05     Completed Date:  1997-12-05     Revised Date:  2007-07-23    
Medline Journal Info:
Nlm Unique ID:  0107600     Medline TA:  Eur J Biochem     Country:  GERMANY    
Other Details:
Languages:  eng     Pagination:  195-222     Citation Subset:  IM    
Bijvoet Center, Department of Bio-Organic Chemistry, Utrecht University, The Netherlands.
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MeSH Terms
Carbohydrate Conformation
Carbohydrate Sequence
Carbohydrates / chemistry
Chromatography, High Pressure Liquid
Helix (Snails) / chemistry*
Hemocyanin / chemistry*
Magnetic Resonance Spectroscopy
Molecular Sequence Data
Molecular Structure
Oligosaccharides / chemistry,  isolation & purification
Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
Polysaccharides / chemistry
Reg. No./Substance:
0/Carbohydrates; 0/Oligosaccharides; 0/Polysaccharides; 9013-72-3/Hemocyanin; EC 3.5.-/Amidohydrolases; EC Asparagine Amidase

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