Structure of the glycan chain from the surface layer glycoprotein of Clostridium thermophydrosulfuricum L77-66

• Published in: Biochimica et biophysica acta. General subjects Vol. 1117; no. 1; pp. 71 - 77
• Main Authors: Altman, Eleonora, Brisson, Jean-Robert, Gagné, Stéphane M., Kolbe, Judith, Messner, Paul, Sleytr, Uwe B.
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 21.07.1992
• Subjects: COSY; Glycan; Glycan structure; NOESY; S-layer; Surface layer glycoprotein; TPPI; NEOSY; HMQC; HMBC; S-layer; GARP; NOESY; Surface layer glycoprotein; Glycan structure; COSY; Glycan; HOHAHA
• ISSN: 0304-4165; 1872-8006
• DOI: 10.1016/0304-4165(92)90164-P

The thermohilic eubacterium Clostridium thermohydrosulfuricum L77-66 is covered by a crystalline surface layer composed of identical glyxoprotein subunits which are arranged in a hexagonal lattice with centre-to-center spacings of approx. 14.3 nm. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis of cell wall preparations showed the presence of several broadened, carbohydrate-containing bands in a molecular mass of 90 to 200 kDa. A total carbohydrate content of approx. 14% was determined in the purified surface layer glycoprotein. Chemical deglycosylation of this material by triflouromethamesulfonic acid resulted in the disappearance of the complex banding pattern. Only a single band with a molecular mass 82 kDa remained visible upon Coomassie staining. After proteolytic digestion of the surface layer glycoprotein a single glycopeptide fraction with an apparent molecular mass of approx. 25 kDa was obtained by gel filtration. Composition analysis, nethylation, periodate oxidation and a combination of homonuclear and 1H-detected heteronuclear shift-correlated nuclear magnetic resonance experiments established the following structure for the glycan chain layer glycoprotei: ▪