title: Characterization of the glycan structure of a major glycopeptide from the surface layer glycoprotein of Clostridium thermosaccharolyticum E207-71.
abstract: The squarely arranged surface layer (S-layer) glycoprotein of Clostridium thermosaccharolyticum E207-71 was isolated from bacterial cells which were grown under defined culture conditions. By sodium dodecyl sulfate polyacrylamide gel electrophoresis, the S-layer showed a series of distinct bands with apparent molecular masses in the range 83-210 kDa. Upon deglycosylation by trifluoromethanesulfonic acid, only the single band at 83 kDa remained unchanged. After pronase digestion of the intact S-layer glycoprotein, the degradation products were isolated by gel-permeation chromatography, cation-exchange chromatography and isoelectric focusing. Three main fractions and an amino sugar containing minor fraction were obtained. The main fractions, which showed identical carbohydrate compositions, were further purified by reverse-phase chromatography and characterized by monosaccharide analysis, Smith degradation, methylation analysis, and one-dimensional and two-dimensional nuclear magnetic resonance spectroscopy. The combined chemical and spectroscopical evidence suggest the following glycan structure for the main fractions: [Sequence: See text]
authors.affiliation: Institute for Biological Sciences, National Research Council, Ottawa, Canada.
authors.forename: E
authors.initials: E
authors.lastname: Altman
authors.forename: C
authors.initials: C
authors.lastname: Schäffer
authors.forename: J R
authors.initials: JR
authors.lastname: Brisson
authors.forename: P
authors.initials: P
authors.lastname: Messner
mesh_terms: D000595:Amino Acid Sequence; D002240:Carbohydrate Sequence; D003013:Clostridium / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D008562:Membrane Glycoproteins / Q000737:chemistry*; D008969:Molecular Sequence Data; D015394:Molecular Structure; D011134:Polysaccharides / Q000737:chemistry*
chemical_list: D008562:Membrane Glycoproteins; D011134:Polysaccharides; C055764:S-layer proteins