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T. Wugeditsch, N. Zachara, M. Puchberger, P. Kosma, A. Gooley, P. Messner:
"Structural heterogeneity in the core oligosaccharide of the S-layer glycoprotein from Aneurinibacillus thermoaerophilus DSM 10155";
Glycobiology, 9 (1999), 8; 787 - 795.

The surface layer glycoprotein of Aneurinibacilllls thermoaerophilus DSM 10155 has a total carbohydrate content of 15% (by mass), consisting of O-linked oligosaccharide chains. After proteolytic digestion of the S-Iayer glycoprotein by Pronase 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)-α-L-Rhap-(1→3)-ß-D-glycero-D-manno-Hepp(1→ (Kosma et al., 1995b, Glycobiology, 5, 791-796) were obtained. Combined evidence from modified Edmandegradation 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 a-L-Rhap-(1 →3)-α-L-Rhap-(1 →3)-ß-D-GaIpNAc-(1 →O)-Thr/Ser and the truncated forms a-L-Rhap-(1 →3)-ß-D-GaIpNAc(1 →O)-Thr/Ser and ß-D-GaIpNAc-(1 →O)-Thr/Ser. All glycopeptides possessed the novel linkage types ß-D-GaIpNAc(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 ofthe mass spectroscopy experiments. Based on the S-Iayer glycopeptide structure, a pathway for its biosynthesis is suggested.

core oligosaccharide / eubacterialglycoprotein / heterogeneity / surface-Iayer (S-layer)

http://dx.doi.org/10.1093/glycob/9.8.787