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
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
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.