THE STRUCTURES OF OLIGOSACCHARIDE BISPHOSPHATES ISOLATED FROM THE LIPOPOLYSACCHARIDE OF A RECOMBINANT ESCHERICHIA-COLI STRAIN EXPRESSING THE GENE GSEA [3-DEOXY-D-MANNO-OCTULOPYRANOSONIC ACID (KDO) TRANSFERASE]OF CHLAMYDIA-PSITTACI 6BC

Citation
O. Holst et al., THE STRUCTURES OF OLIGOSACCHARIDE BISPHOSPHATES ISOLATED FROM THE LIPOPOLYSACCHARIDE OF A RECOMBINANT ESCHERICHIA-COLI STRAIN EXPRESSING THE GENE GSEA [3-DEOXY-D-MANNO-OCTULOPYRANOSONIC ACID (KDO) TRANSFERASE]OF CHLAMYDIA-PSITTACI 6BC, European journal of biochemistry, 229(1), 1995, pp. 194-200
Citations number
19
Categorie Soggetti
Biology
ISSN journal
00142956
Volume
229
Issue
1
Year of publication
1995
Pages
194 - 200
Database
ISI
SICI code
0014-2956(1995)229:1<194:TSOOBI>2.0.ZU;2-A
Abstract
The lipopolysaccharide from the recombinant strain Escherichia coli F5 15-140 containing the cloned gene gseA [3-deoxy-D-manno-octulopyranoso nic acid (Kdo) transferase] from Chlamydia psittaci 6BC was isolated a nd sequentially de-O-acylated and de-N-acylated. The products were sep arated by high-performance anion-exchange chromatography into three fr actions, two of which contained a single compound. Their structures we re elucidated by high-field NMR spectroscopy as alpha-Kdo-(2-->6)-beta -D-GlcN-(1-->6)-alpha-D-GlcN 1,4'-P-2 (compound 1) (tetrasaccharide bi sphosphate) [Holst, O., Broer, W., Thomas-Gates, J. E., Mamat, U. and Brade, H. (1993) Eur. J. Biochem. 214, 703-710] and alpha-Kdo-(2-->4)- [alpha-Kdo-(2--> alpha-Kdo-(2-->6)-beta-D-GlcN-(1-->6)-alpha-D-GlcN 1, 4'-P-2 (compound 4) (hexasaccharide bisphosphate). The third fraction comprised two pentasaccharide bisphosphates, which could be separated by affinity chromatography using an immobilized monoclonal antibody sp ecific for the trisaccharide alpha-Kdo-(2-->8)-alpha-Kdo-(2-->4)-alpha -Kdo. The bound fraction was identified as alpha-Kdo-(2-->8)-alpha-Kdo -(2-->4)-alpha-Kdo- (2-->6)-beta-D-GlcN-(1-->6)-alpha-D-GlcN 1,4'-P-2 (compound 2) [Hoist, O., Broer, W., Thomas-Gates, J. E., Mamat, U. and Brade, H. (1993) Eur. J. Biochem. 214, 703-710], whereas the unbound fraction was identified as alpha-Kdo-(2-->6)-beta-D-GlcN-(1-->6)-alpha -D-GlcN 1,4'-P-2 (compound 3). This novel Kdo tetrasaccharide extends our knowledge on multifunctional Kdo transferases.