Primary structure of the O-glycosidically linked glycan chain of the crystalline surface layer glycoprotein of Thermoanaerobacter thermohydrosulfuricus L111-69. Galactosyl tyrosine as a novel linkage unit.
The products of Pronase digestion of the crystalline surface layer (S-layer) glycoproteins of Thermoanaerobacter thermohydrosulfuricus strains L111-69 and L110-69 were isolated by gel permeation chromatography, cation exchange chromatography, chromatofocusing, and reversed phase high performance liquid chromatography. Four compounds were obtained which were analyzed by monosaccharide analysis, one- and two-dimensional 500 and 600 MHz 1H and 13C NMR spectroscopy, methylation analysis, gas-liquid chromatography/mass spectrometry, and matrix-assisted laser desorption ionization mass spectrometry. For all glycopeptides we propose the following glycan structure with galactose as the linkage sugar. [formula: see text] The isolated glycopeptides resulted from Pronase cleavage at the glycosylated tyrosine residues. Tyrosine was found as the linkage amino acid in all fractions but the remaining amino acid sequences varied, indicating the presence of different glycosylation sites in the intact S-layer glycoprotein.
Bock
K
K
Department of Chemistry, Carlsberg Laboratory, Valby, Copenhagen, Denmark.
Schuster-Kolbe
J
J
Altman
E
E
Allmaier
G
G
Stahl
B
B
Christian
R
R
Sleytr
U B
UB
Messner
P
P
D001426:Bacterial Proteins / Q000737:chemistry*; D002240:Carbohydrate Sequence; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003013:Clostridium / Q000737:chemistry*; D006020:Glycopeptides / Q000737:chemistry; D006868:Hydrolysis; D013058:Mass Spectrometry; D008745:Methylation; D008969:Molecular Sequence Data; D011134:Polysaccharides / Q000737:chemistry*; D014443:Tyrosine / Q000737:chemistry*
D001426:Bacterial Proteins; D006020:Glycopeptides; D011134:Polysaccharides; D014443:Tyrosine