The structure of the carbohydrate backbone of the lipopolysaccharide from Acinetobacter baumannii strain ATCC 19606 (original) (raw)
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Carbohydrate Research, 2001
Lipopolysaccharides (LPSs) of Chlamydophila psittaci 6BC and Chlamydophila pneumoniae Kajaani 6 contain 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo), GlcN, organic bound phosphate, and fatty acids in the molar ratios of 3:2:2.2:4.8 and 2.9:2:2.1:4.9, respectively. The LPSs were immunoreactive with a monoclonal antibody against a family-specific epitope of chlamydial LPS. This finding, together with methylation analyses of both LPSs and MALDI-TOF MS experiments on de-O-, and de-O,N-acylated LPSs, indicate the presence of a Kdo trisaccharide proximal to lipid A having a structure a-Kdo-(2 8)-a-Kdo-(2 4)-a-Kdo, which appears to be the main component of the core region in the native chlamydial LPSs. In the de-O-acylated LPSs from Chl. psittaci 6BC and Chl. pneumoniae Kajaani 6, two major molecular species are present that differ in distribution of amide-bound hydroxy fatty acids over both GlcN. It appears that either two (R)-3-hydroxy-18-methylicosanoic acids or one (R)-3-hydroxy-18-methylicosanoic acid and one (R)-3-hydroxyicosanoic acid are attached to the GlcN residues. In contrast, the de-O-acylated LPS of Chl. psittaci PK 5082 contains one major molecular species that has two (R)-3-hydroxyicosanoic acid residues attached to two GlcN residues.
Journal of Biological Chemistry, 1998
For the first time, the tetrasaccharide Kdo␣235Kdo␣235(Kdo␣234)Kdo (Kdo is 3-deoxy-Dmanno-oct-2-ulopyranosonic acid) has been identified in a bacterial lipopolysaccharide (LPS), i.e. in the core region of LPS from Acinetobacter baumannii NCTC 10303. The LPS was analyzed using compositional analysis, mass spectrometry, and NMR spectroscopy. The disaccharide D-GlcpN136D-GlcpN, phosphorylated at O-1 and O-4, was identified as the carbohydrate backbone of the lipid A. The Kdo tetrasaccharide is attached to O-6 of this disaccharide and is further substituted by short L-rhamnoglycans of varying length and by the disaccharide D-GlcpNAc␣134D-GlcpNA (GlcpNA, 2-amino-2-deoxy-glucopyranosuronic acid). The core region is not substituted by phosphate residues and represents a novel core type of bacterial LPS. The complete carbohydrate backbone of the LPS is shown in Structure I as follows:
European Journal of Biochemistry, 1995
The lipopolysaccharide from the recombinant strain Escherichia coli F515-140 containing the cloned gene gseA [3-deoxy-D-manno-octu~opyranosonic acid (Kdo) transferase] from Chlamydia psittaci 6BC was isolated and sequentially de-0-acylated and deN -acylated. The products were separated by highperformance anion-exchange chromatography into three fractions, two of which contained a single compound. Their structures were elucidated by high-field NMR spectroscopy as a-Kdo-(2+4)-a-Kdo-(2+6)-P-~-GlcN-(1+6)-aD -GlcN 1,4'-P, (compound 1) (tetrasaccharide bisphosphate) [Holst, O., Broer, W., Thomas-Oates, J. E., Mamat, U. & Brade, H. (1993) Eur. J. Biochem. 214, 703-7101 and a-Kdo-(2+4)-[a-Kdo-(2-+8)-]-a-Kdo-(2-t4)-a-Kdo-(2+6)-~-~-GlcN-(1-t6)-a-~-GlcN 1,4'-P2 (compound 4) (hexasaccharide bisphosphate). The third fraction comprised two pentasaccharide bisphosphates, which could be separated by affinity chromatography using an immobilized monoclonal antibody specific for the trisaccharide a-Kdo-(2+8)-a-Kdo-(2-.4)-a-Kdo. The bound fraction was identified as a-Kdo-(2-+8)-a-Kdo-(2+4)-a-Kdo-(2-+6)-~-~-GlcN-(1+6)-a-~-GlcN 1,4'-P, (compound 2) [Holst, O., Broer, W., Thomas-Oates, J. E., Mamat, U. & Brade, H. (1993) Eur J. Biochem. 214, 703-7101, whereas the unbound fraction was identified as a-Kdo-(2-.4)-cx-Kdo-(2-.4)-a-Kdo-(2+6)-~-~-GlcN-(1+6)-a-~-GlcN 1,4'-P, (compound 3). This novel Kdo tetrasaccharide extends our knowledge on multifunctional Kdo transferases.
Journal of Biological Chemistry, 1998
In a preliminary investigation (Sü sskind, M., Mü ller-Loennies, S., Nimmich, W., Brade, H., and Holst, O. (1995) Carbohydr. Res. 269, C1-C7), we identified after deacylation of lipopolysaccharides (LPS) from Klebsiella pneumoniae ssp. pneumoniae rough strain R20 (O1 ؊ :K20 ؊) as a major fraction the oligosaccharide, GalpA1 3 6Glcp1 3 4Hepp␣1 3 5Kdo␣2 3 6GlcpN1 3 6GlcpN␣1 3 P 3 4 4 1 1 1 threo-hex-4-enuronop1 3 3Hepp␣1 Kdo␣2 P 7 (L-glycero-D-manno-heptopyranose; L,D-Hepp), where all hexoses possess the D-configuration. Sugars marked with an asterisk are present in nonstoichiometric amounts. The structure is unique with regard to the presence of an ␣132-linked D-glycero-D-manno-heptoglycan (oligosaccharide), which has not been described to date, and does not contain phosphate substituents in the core region. Fatty acid analysis of lipid A identified (R)-3-hydroxytetradecanoic acid as sole amide-linked fatty acid and (R)-3-hydroxytetradecanoic acid, tetradecanoic acid, small amounts of 2-hydroxytetradecanoic acid, hexadecanoic acid, and traces of dodecanoic acid as ester-linked fatty acids, substituting the carbohydrate backbone D-GlcpN4P136D-GlcpN␣1P. The nonreducing GlcN carries four fatty acids, present as two 3-O-tetradecanoyltetradecanoic acid residues, one of which is amide-linked and the other ester-linked to O-3. The reducing GlcN is substituted in a nature fraction of lipid A by two residues of (R)-3hydroxytetradecanoic acid, one in amide and the other in ester linkage at O-3. Two minor fractions of lipid A were identified; in one, the amide-linked (R)-3-hydroxytetradecanoic acid at the reducing GlcN is esterified with hexadecanoic acid, resulting in 3-O-hexadecanoyltetradecanoic acid, and in the second, one of the 3-O-tetradecanoyltetradecanoic acid residues at the nonreducing GlcN is replaced by 3-O-dodecanoyltetradecanoic acid. Thus, the complete structure of LPS is as shown in Fig. 1. After immunization of BALB/c mice, two monoclonal antibodies were obtained that were shown to be specific for the core of LPS from K. pneumoniae ssp. pneumoniae, since they did not react with LPS or whole-cell lysates of a variety of other Gram-negative species. Both monoclonal antibodies could be inhibited by LPS but not by isolated oligosaccharides and are thus considered to recognize a conformational epitope in the core region.
European Journal of Biochemistry
The structure of the lipopolysaccharide (LPS) from Acinetobacter haemoZyticus strain NCTC 10305 (DNA group 4) was elucidated by means of analytical chemistry, NMK spectroscopy and fast-atombombardment mass spectrometry. Several oligosaccharides were obtained after deacylation or successive de-0-acylation, dephosphorylation, reduction, and de-N-acylation of LPS. In the major fraction of the LPS, the core is attached to the lipid A through D-g~ycero-D-tu~o-2-octulopyranosonic acid (KO), whereas in a minor fraction (< 20%) KO is replaced by 3-deoxy-D-manno-octulopyranosonic acid (Kdo). The structures of the phosphorylated carbohydrate backbones of these LPS fractions are a-IKdo 2 4 I a-Dha-(2-6)-/-Glc-( 1 -4)-u-Dha-(2~6)-u-GIc-(I -6)-n-Glc-(l-6)-~-G1~-(1-4)-~-Glc6P-(1-5)-u-Sug-(2~6)-/-GlcN4P-(1-6)-~-GlcN1 P