The Surface Layer (S-layer) Glycoprotein of Geobacillus stearothermophilus NRS 2004/3a

Characterization of methyl sugars, 3-deoxysugars and methyl deoxysugars in marine high molecular weight dissolved organic matter

“…We found two anomeric protons at 5.32 ppm (H-1, α-anomer; d, J 1,2 =1.6 Hz) and 4.92 ppm (H-1, β-anomer; broad singlet), H-C-OH protons at 3.39-3.87 ppm, one O-methyl group at 3.51 ppm, and two methyl groups at 1.31 ppm (J 5,6 = 5.3 Hz) and 1.33 ppm (J 5,6 = 6.7 Hz) for the α and β anomers respectively. We found good agreement between our spectral data and 2-Omethylrhamnose as the terminal sugar in a glycan polymer (Schäffer et al, 2002).…”

“…In the HSQC spectrum we observed a correlation between βH-3 (3.37 ppm) and a carbon at 82.4 ppm, which we assigned as βC-3, further establishing the site of methylation. There is a cross peak between H-4 (3.45 ppm) and a carbon at 71.4 ppm, which we assigned as βC-4, in good agreement with the values assigned by Popper and Fry (2003) We used the same reasoning to identify F 3 B2 as a α/ß mixture of 2-Omethylrhamnose (XVI) (Zdorovenko et al 2001;Schäffer et al 2002). Fractions F 3 B1…”

Recombinant Glycans on an S‐Layer Self‐Assembly Protein: A New Dimension for Nanopatterned Biomaterials

“…For the introduction of a distinctive protein N-glycosylation consensus sequence from the C. jejuni AcrA N-glycoprotein replacing the naturally O-glycosylation sites of the S-layer protein, [19,20] a modified sgsE gene containing an artificial KpnI site at position nt 1830 (corresponding to amino acid position 611), coding for a protein with a PelB signal peptide and lacking the amino acid sequence between L 610 and E 804 was constructed. N-and Cterminal parts of sgsE were amplified separately by PCR using the primer pairs A_sgsE_for(NcoI)/sgsEteil_rev(KpnI) and sgsE-teil_for(KpnI)/sgsE_rev(XhoI), respectively.…”

“…SgsE is a 903-amino acid protein, which includes a 30-amino acid signal peptide aligned by an entropy-driven process into a 2D crystalline array [8] with oblique (p2) symmetry exhibiting nanometer-scaled periodicity (lattice parameters: a = 11.6 nm, b = 9.4 nm, γ ≈ 78°). [18,19] SgsE was chosen for proof of concept because it is naturally O-glycosylated with long-chain poly-L-rhamnans linked via a β-D-galactose residue to the amino acids threonine 590 , threonine 620 , and serine 794 of the S-layer protein. [19,20] The protein-inherent glycosylation sites are predicted to form a loop structure that is spatially accessible to the glycosylation machinery of the bacterium.…”

“…[18,19] SgsE was chosen for proof of concept because it is naturally O-glycosylated with long-chain poly-L-rhamnans linked via a β-D-galactose residue to the amino acids threonine 590 , threonine 620 , and serine 794 of the S-layer protein. [19,20] The protein-inherent glycosylation sites are predicted to form a loop structure that is spatially accessible to the glycosylation machinery of the bacterium. Thus, these sites offer ideal targets for engineering of specific glycosylation sequences that would be recognized by heterologous oligosaccharyl:protein transferases, as required for S-layer neoglycoprotein design.…”