Structural heterogeneity in the core oligosaccharide of the S-layer glycoprotein from Aneurinibacillus thermoaerophilus DSM 10155

Glycobiology. 1999 Aug;9(8):787-95. doi: 10.1093/glycob/9.8.787.


The surface layer glycoprotein of Aneurinibacillus thermoaerophilus DSM 10155 has a total carbohydrate content of 15% (by mass), consisting of O-linked oligosaccharide chains. After proteolytic digestion of the S-layer glycoprotein byPronase E and subsequent purification of the digestion products by gel permeation chromatography, chromatofocusing and high-performance liquid chromatography two glycopeptide pools A and B with identical glycans and the repeating unit structure -->4)-alpha-l-Rha p -(1-->3)-beta-d- glycero -d- manno -Hep p -(1--> (Kosma et al., 1995b, Glycobiology, 5, 791-796) were obtained. Combined evidence from modified Edman-degradation in combination with liquid chromatography electrospray mass-spectrometry and nuclear magnetic resonance spectroscopy revealed that both glycopeptides contain equal amounts of the complete core structure alpha-l-Rha p -(1-->3)-alpha-l-Rha p -(1-->3)-beta-d-Gal p NAc-(1-->O)-Thr/Ser and the truncated forms alpha-l-Rha p -(1-->3)-beta-d-Gal p NAc-(1-->O)-Thr/Ser and beta-d-Gal p NAc-(1-->O)-Thr/Ser. All glycopeptides possessed the novel linkage types beta-d-Gal p NAc-(1-->O)-Thr/Ser. The different cores were substituted with varying numbers of disaccharide repeating units. By 300 MHz proton nuclear magnetic resonance spectroscopy the complete carbohydrate core structure of the fluorescently labeled glyco-peptide B was determined after Smith-degradation of its glycan chain. The NMR data confirmed and complemented the results of the mass spectroscopy experiments. Based on the S-layer glycopeptide structure, a pathway for its biosynthesis is suggested.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacillus / chemistry*
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / isolation & purification
  • Carbohydrate Conformation
  • Carbohydrate Sequence
  • Chromatography, Gel
  • Chromatography, High Pressure Liquid
  • Glycopeptides / chemistry
  • Glycopeptides / isolation & purification
  • Membrane Glycoproteins / chemistry*
  • Membrane Glycoproteins / isolation & purification
  • Molecular Sequence Data
  • Nuclear Magnetic Resonance, Biomolecular
  • Oligosaccharides / chemistry*
  • Oligosaccharides / isolation & purification
  • Pronase
  • Spectrometry, Mass, Secondary Ion


  • Bacterial Proteins
  • Glycopeptides
  • Membrane Glycoproteins
  • Oligosaccharides
  • S-layer proteins
  • Pronase