Analysis of Binding Sites of Sugar Chains by Methylation Analysis

  • Sumihiro Hase


The linkage structures between the monomer units in most proteins and nucleic acids are one of a kind; however, for carbohydrates the linkage structures between the monosaccharide units, acetal or ketal linkage (glycosidic linkage), are more than one of a kind, including the anomeric configuration and the linkage positions of available OH groups. Therefore, determination of the linkage positions of the monosaccharide residues is inevitable for determination of the chemical structures of sugar chains. This analysis may be partly done by Smith degradation, mass spectrometry, hydrolysis with specific enzymes, and NMR; however, methylation analysis is the most reliable method in this field especially for determination of novel sugar structures.


Methylation Analysis Glycosidic Linkage Linkage Structure Sugar Chain Monosaccharide Residue 


  1. Blakeney AB, Stone BA (1985) Methylation of carbohydrates with lithium methylsulphinyl carbanion. Carbohydr Res 140:319–324CrossRefGoogle Scholar
  2. Ciucanu I, Kerek F (1984) A simple and rapid method for the permethylation of carbohydrates. Carbohydr Res 131:209–217CrossRefGoogle Scholar
  3. Geyer R, Geyer H, Kühnhardt S, Mink W, Stirm S (1983) Methylation analysis of complex carbohydrates in small amounts: capillary gas chromatography-mass fragmentography of methylalditol acetates obtained from N-glycosidically linked glycoprotein oligosaccharides. Anal Biochem 133:197–207PubMedCrossRefGoogle Scholar
  4. Geyer R, Geyer H (1994) Saccharide linkage analysis using methylation and other techniques. In: Lennarz WJ, Hart GW (eds) Methods in enzymology, vol 230. Academic Press, San Diego, pp 86–108Google Scholar
  5. Hakomori S (1964) A rapid permethylation of glycolipid, and polysaccharide catalyzed by methylsulfinyl carbanion in dimethyl sulfoxide. J Biochem 55:205–208PubMedGoogle Scholar
  6. Parente JP, Cardon P, Leroy Y, Montreuil J, Fournet B (1985) A convenient method for methylation of glycoprotein glycans in small amounts by using lithium methylsulfinyl carbanion. Carbohydr Res 141:41–47PubMedCrossRefGoogle Scholar
  7. Wakabayashi H, Natsuka S, Mega T, Otsuki N, Isaji M, Naotsuka M, Koyama S, Kanamori T, Sakai K, Hase S (1999) Novel proteoglycan linkage tetrasaccharides of human urinary soluble thrombomodulin, SO4-3GlcAβ1-3Galβ1-3(±Siaα2-6)Galβ1-4Xyl. J Biol Chem 274:5436–5442PubMedCrossRefGoogle Scholar

Copyright information

© Springer 2008

Authors and Affiliations

  • Sumihiro Hase
    • 1
  1. 1.Department of Chemistry, Graduate School of ScienceOsaka UniversityToyonaka, OsakaJapan

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