Complete structure of the tyrosine-linked saccharide moiety from the surface layer glycoprotein of Clostridium thermohydrosulfuricum S102-70

Abstract

In this study, we have extended and completed a previous investigation (P. Messner, R. Christian, J. Kolbe, G. Schulz, and U. B. Sleytr, J. Bacteriol. 174:2236Bacteriol. 174: -2240Bacteriol. 174: , 1992) in which we demonstrated for the first time in prokaryotic organisms the presence of a novel 0-glycosidic linkage via tyrosine. The surface layer glycoprotein of the eubacterium Clostridium thermohydrosufuricum S102-70 is arranged in a hexagonal lattice, with center-to-center spacings of approximately 16.3 nm. Molecular weight determination by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of both glycosylated and chemically deglycosylated surface layer glycoprotein showed values for the monomeric subunits of 94,000 and 87,500, respectively. Glycopeptide fractions obtained after exhaustive pronase digestion of purified, intact glycoprotein were isolated by reversed-phase liquid chromatography. One-and two-dimensional nuclear magnetic resonance studies, together with chemical analyses and plasma desorption time-of-flight mass spectrometry, were used to elucidate the structure of the hexasaccharide moiety linked by the novel 0-glycosidic linkage to tyrosine. The combined evidence suggests the following structure:

New Insights into the Glycosylation of the Surface Layer Protein SgsE from Geobacillus stearothermophilus NRS 2004/3a

“…However, the apparent molecular masses of S-layer proteins with extended glycan chains are usually too high (16,24,39). The finding that the masses of nonglycosylated or chemically deglycosylated S-layer proteins correspond well with the theoretical values from protein sequencing experiments indicates that glycosylation of these proteins is responsible for the aberrant migration behavior of bacterial Slayer glycoproteins on SDS-PA gels (3,6,35). Upon chemical deglycosylation of the S-layer glycoprotein of G. stearothermophilus NRS 2004/3a, a single 93-kDa band representing the nonglycosylated SgsE protein remains visible on the gel (20,35).…”

Crystalline Bacterial Cell Surface Layers (S Layers): From Supramolecular Cell Structure to Biomimetics and Nanotechnology

“…Thermoanaerobacter thermohydrosulfuricus S102-70 (formerly Clostridium thermohydrosulfuricum) [77,78] b…”

UDP-galactofuranose Precursor Required for Formation of the Lipopolysaccharide O Antigen of Klebsiella pneumoniae Serotype O1 Is Synthesized by the Product of the rfbDKPO1 Gene

“…16 -20) contain Galf. Galactofuranosyl residues are a central component in the mycolyl-arabinogalactan complex, which is characteristic of the cell walls of mycobacteria (21) and the related genera Nocardia and Rhodococcus (22), in the lipoglycan of Mycoplasma mycoides (23), in the paracrystalline S-layer glycoprotein of Clostridium thermohydrosulfuricum S102-70 (24), and in the cellulosome glycoproteins of Clostridium thermocellum (25) and Bacteroides cellulosolvens (26). In eukaryotes, galactofuranosyl residues are found in the lipophosphoglycan of Leishmania donovani (27), Leishmania major (28), and Leishmania mexicana (29), in the N-linked glycoproteins of Crithidia spp.…”