Taxonomic group: bacteria / Firmicutes
(Phylum: Firmicutes)
The structure was elucidated in this paperNCBI PubMed ID: 8444787Journal NLM ID: 2985120RPublisher: American Society for Microbiology
Institutions: Zentrum für Ultrastrukturforschung, Universität für Bodenkultur, Vienna, Austria
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. 174:2236-2240, 1992) in which we demonstrated for the first time in prokaryotic organisms the presence of a novel O-glycosidic linkage via tyrosine. The surface layer glycoprotein of the eubacterium Clostridium thermohydrosulfuricum 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 O-glycosidic linkage to tyrosine. The combined evidence suggests the following structure: β-D-Galf-(1→3)-α-D-Galp- (1→2)-α-L-Rhap-(1→3)-α-D-Manp-(1→3)-α-L-Rhap-(1→3)-β- D-Glcp-(1→4)-L-Tyr.
structure, mass spectrometry, Clostridium, glycoprotein, tyrosine
Structure type: oligomer ; 87500-94000
Location inside paper: abstract, p.1254, GP III
Trivial name: S-layer glycan
Compound class: glycopeptide
Contained glycoepitopes: IEDB_130701,IEDB_136095,IEDB_136105,IEDB_136906,IEDB_137472,IEDB_141794,IEDB_142488,IEDB_144983,IEDB_146664,IEDB_151528,IEDB_152206,IEDB_189517,IEDB_190606,IEDB_225177,IEDB_885823,IEDB_983930,IEDB_983931,SB_192,SB_44,SB_67,SB_7,SB_72
Methods: 13C NMR, 1H NMR, NMR-2D, SDS-PAGE, chemical analysis, PD-MS, NMR-1D, electron microscopy, deglycosylation
NCBI Taxonomy refs (TaxIDs): 1485
Show glycosyltransferases
NMR conditions: in D2O at 310 K
[as TSV]
13C NMR data:
Linkage Residue C1 C2 C3 C4 C5 C6 C7 C8 C9
7,3,3,3,2,3 bDGalf 109.97 82.38 77.71 83.67 71.74 63.61
7,3,3,3,2 aDGalp 98.61 68.25 77.84 70.09 71.68 61.73
7,3,3,3 aLRhap 94.97 77.28 70.59 72.84 70.00 17.43
7,3,3 aDManp 97.11 67.53 76.32 65.71 73.60 61.73
7,3 aLRhap 101.82 67.28 75.34 71.16 69.85 17.55
7 bDGlcp 100.90 74.41 82.93 68.80 77.02 61.45
xLTyr ? 56.94 35.76 117.99 130.69 131.69 156.92 131.69 130.69
1H NMR data:
Linkage Residue H1 H2 H3 H4 H5 H6 H7 H8 H9
7,3,3,3,2,3 bDGalf 5.249 4.213 4.090 4.068 3.854 3.672-3.721
7,3,3,3,2 aDGalp 5.121 3.976 4.027 4.183 4.305 3.746-3.746
7,3,3,3 aLRhap 5.150 4.092 3.994 3.580 4.026 1.318
7,3,3 aDManp 5.113 4.233 4.012 3.817 3.980 3.823-3.901
7,3 aLRhap 5.239 4.324 3.943 3.569 4.095 4.301
7 bDGlcp 5.146 3.705 3.732 3.589 3.654 3.773-3.948
xLTyr - 4.085 3.105-3.252 - 7.294 7.135 - 7.135 7.294
1H/13C HSQC data:
Linkage Residue C1/H1 C2/H2 C3/H3 C4/H4 C5/H5 C6/H6 C7/H7 C8/H8 C9/H9
7,3,3,3,2,3 bDGalf 109.97/5.249 82.38/4.213 77.71/4.090 83.67/4.068 71.74/3.854 63.61/3.672-3.721
7,3,3,3,2 aDGalp 98.61/5.121 68.25/3.976 77.84/4.027 70.09/4.183 71.68/4.305 61.73/3.746-3.746
7,3,3,3 aLRhap 94.97/5.150 77.28/4.092 70.59/3.994 72.84/3.580 70.00/4.026 17.43/1.318
7,3,3 aDManp 97.11/5.113 67.53/4.233 76.32/4.012 65.71/3.817 73.60/3.980 61.73/3.823-3.901
7,3 aLRhap 101.82/5.239 67.28/4.324 75.34/3.943 71.16/3.569 69.85/4.095 17.55/4.301
7 bDGlcp 100.90/5.146 74.41/3.705 82.93/3.732 68.80/3.589 77.02/3.654 61.45/3.773-3.948
xLTyr 56.94/4.085 35.76/3.105-3.252 130.69/7.294 131.69/7.135 131.69/7.135 130.69/7.294
1H NMR data:
| Linkage | Residue | H1 | H2 | H3 | H4 | H5 | H6 | H7 | H8 | H9 |
|---|
| 7,3,3,3,2,3 | bDGalf | 5.249 | 4.213 | 4.090 | 4.068 | 3.854 | 3.672
3.721 | |
| 7,3,3,3,2 | aDGalp | 5.121 | 3.976 | 4.027 | 4.183 | 4.305 | 3.746
3.746 | |
| 7,3,3,3 | aLRhap | 5.150 | 4.092 | 3.994 | 3.580 | 4.026 | 1.318 | |
| 7,3,3 | aDManp | 5.113 | 4.233 | 4.012 | 3.817 | 3.980 | 3.823
3.901 | |
| 7,3 | aLRhap | 5.239 | 4.324 | 3.943 | 3.569 | 4.095 | 4.301 | |
| 7 | bDGlcp | 5.146 | 3.705 | 3.732 | 3.589 | 3.654 | 3.773
3.948 | |
| | xLTyr |
| 4.085 | 3.105
3.252 |
| 7.294 | 7.135 |
| 7.135 | 7.294 |
|
13C NMR data:
| Linkage | Residue | C1 | C2 | C3 | C4 | C5 | C6 | C7 | C8 | C9 |
|---|
| 7,3,3,3,2,3 | bDGalf | 109.97 | 82.38 | 77.71 | 83.67 | 71.74 | 63.61 | |
| 7,3,3,3,2 | aDGalp | 98.61 | 68.25 | 77.84 | 70.09 | 71.68 | 61.73 | |
| 7,3,3,3 | aLRhap | 94.97 | 77.28 | 70.59 | 72.84 | 70.00 | 17.43 | |
| 7,3,3 | aDManp | 97.11 | 67.53 | 76.32 | 65.71 | 73.60 | 61.73 | |
| 7,3 | aLRhap | 101.82 | 67.28 | 75.34 | 71.16 | 69.85 | 17.55 | |
| 7 | bDGlcp | 100.90 | 74.41 | 82.93 | 68.80 | 77.02 | 61.45 | |
| | xLTyr | ? | 56.94 | 35.76 | 117.99 | 130.69 | 131.69 | 156.92 | 131.69 | 130.69 |
|
The spectrum also has 1 signal at unknown position (not plotted). |
There is only one chemically distinct structure:
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Found 1 record.
Displayed record 1
