Structural characterization of the O-polysaccharide of the lipopolysaccharide produced by Salmonella milwaukee O:43 (group U) which possesses human blood group B activity.

The O-polysaccharide of the lipopolysaccharide produced by Salmonella milwaukee O:43 (group U) was shown by composition analysis, methylation, periodate oxidation, and 1H and 13C nuclear magnetic resonance spectroscopic analytical methods to be a polymer of branched pentasaccharide repeating units having the structure: [formula: see text] The blood-group activity of the O-polysaccharide was established by its serological reactivity with a specific monoclonal antibody to human blood group B, using passive hemagglutination and ELISA assays, indicating the common antigenic epitope to be a nonreducing terminal trisaccharide unit composed of L-Fucp and D-Galp (1:2) residues.     

Characterisation of the Salmonella O:8 antigen in the O-chain of the lipopolysaccharide produced by Salmonella virginia O:8.

The antigenic O-polysaccharide of the lipopolysaccharide produced by Salmonella virginia (O:8), analyzed by methylation, partial acid hydrolysis, and one- and two-dimensional nuclear magnetic resonance methods, was found to be a polymer of a repeating pentasaccharide unit composed of D-mannose, D-galactose, L-rhamnose, D-abequose, and O-acetyl (2:1:1:1:1.3) and having the following structure: [formula; see text] The disaccharide structure alpha-D-Abep-(1----3)-L-Rhap was identified as the Salmonella O:8 antigenic factor epitope, since the removal of alpha-D-Abep residues from the O-polysaccharide left a residual tetrasaccharide repeating unit backbone that did not show reaction with Salmonella type O:8 factor antiserum.     

Structure of the lipopolysaccharide antigenic O-chain produced by Salmonella livingstone (O:6,7)

The lipopolysaccharide produced by Salmonella livingstone (O:6,7) was composed of an antigenic O-polysaccharide which was shown by composition, methylation analysis, and high resolution nuclear magnetic resonance studies to be a high molecular weight polymer containing D-glucose, 2-acetamido-2-deoxy-D-glucose, and D-mannose residues (1:1:4) composed in a repeating hexasaccharide unit having the structure: (formula; see text)     

Structure of a teichoic acid-like O-polysaccharide of Escherichia coli O29

A teichoic acid-like O-polysaccharide was isolated by mild acid degradation of the lipopolysaccharide (LPS) of Escherichia coli O29. The O-polysaccharide and an oligosaccharide obtained by dephosphorylation of the O-polysaccharide were studied by sugar analysis along with 1H and 13C NMR spectroscopy. The following structure of the branched oligosaccharide repeating unit, containing five monosaccharide residues and glycerol 1-phosphate (D-Gro-1-P), was established: [carbohydrate structure: see text].     

Structure of the O-polysaccharide chain of lipopolysaccharide from Pseudomonas aeruginosa IID 1001 (ATCC 27577)

Structural studies were carried out on an acidic O-polysaccharide released by mild acid treatment from the lipopolysaccharide of Pseudomonas aeruginosa IID 1001 (ATCC 27577), which is serologically related to the serotypes Habs O3, Lanyi O1, and Wokatsch O13 in other serological classifications of Pseudomonas aeruginosa. The composition and data from structural analyses including H-NMR and C-NMR measurements, methylation, and Smith degradation showed that the structure of the IID 1001 O-polysaccharide was coincident with that of the Habs O3 and Lanyi O1 antigens (or Wokatsch O13). However, whereas solvolysis of the O-antigen of Habs O3 as well as that of Lanyi O1 by hydrogen fluoride has been reported to yield a reducing trisaccharide, GlcNAc(alpha 1----4)GalNAcA(alpha 1----3)Bac2NAc4Nacyl (acyl represents a 3-hydroxybutanoyl group), hydrogen fluoride hydrolysis of IID 1001 O-polysaccharide yielded a nonreducing trisaccharide with the reducing terminal bacillosamine residue linked at C-1 to the hydroxyl group of its N-acyl substituent, 3-hydroxybutanoic acid. These results, in combination with mass spectral data, led to the most likely structure for the tetrasaccharide repeating unit of the IID 1001 O-polysaccharide, (formula; see text) in which the location of N-acyl groups on bacillosamine residues differs from that in the O-antigens of Habs O3 and Lanyi O1 (or Wokatsch O13).     

Structural studies on O-specific polysaccharides of lipopolysaccharides from Yersinia enterocolitica serovars O:1,2a,3, O:2a,2b,3 and O:3

Lipopolysaccharides from Yersinia enterocolitica serovars O:1,2a,3, O:2a,2b,3 and O:3 have been isolated and characterized. 6-Deoxy-L-altrose residues were shown to be the main constituents of lipopolysaccharides isolated in addition to residues of L-rhamnose, D-glucose, D-galactose, 2-acetamido-2-deoxy-D-glucose, 2-acetamido-2-deoxy-D-galactose, D-glycero-D-manno-heptose and L-glycero-D-manno-heptose, 3-deoxy-D-manno-octulosonic acid being minor components of sugar chains. Mild hydrolysis of lipopolysaccharides with acetic acid furnished O-specific polysaccharides, which are composed of 6-deoxy-L-altrose. Using 13C-NMR spectroscopy and methylation data, the structural features of backbones have been elucidated as follows: ----2)-6d-L-Altp(beta 1----2)-6d-L-Altp(beta 1----3)-6d-L-Altp)(beta 1----for serovars O:1,2a,3 and O:2a,2b,3;----2)-6d-L-Altp(beta 1----for serovar O:3. In addition, O-polysaccharide of serovar O:2a,2b,3 was found to contain an O-acetyl group at the C-3 position of some 1,2-linked sugar residues.     

The structure of the O-polysaccharide from Pseudomonas aeruginosa IID 1009 (ATCC 27585)

Structural studies were carried out on the O-polysaccharide fraction obtained by mild acid treatment of the lipopolysaccharide from Pseudomonas aeruginosa IID 1009 (ATCC 27585). The O-polysaccharide was composed of L-rhamnose, N-acetyl-D-quinovosamine, and N-acetyl-L-galactosaminuronic acid in a molar ratio of 1:1:1. The results from analysis of fragments obtained by hydrogen fluoride hydrolysis of O-polysaccharide, together with data on methylation analysis and nuclear magnetic resonance spectroscopic analysis, led to the most likely structure of the repeating units of the polymer chain ----4)L-GalNAcA(alpha 1----3)D-QuiNAc(alpha 1----3)L-Rha(alpha 1----, in which about 70% of the rhamnose residues were O-acetylated at C-2. This structure coincides with that of the repeating unit of Lanyi 02 a,b polysaccharides.     

The structure of the O-specific chain of lipopolysaccharide from Pseudomonas aeruginosa IID 1012 (ATCC 27588)

Structural studies were carried out on the O-polysaccharide fraction obtained from the lipopolysaccharide of Pseudomonas aeruginosa IID 1012, the standard strain of Homma serogroup K, by mild acid treatment. The O-polysaccharide was composed of L-rhamnose, N-acetyl-D-quinovosamine, and N-acetyl-D-galactosaminuronic acid. The results from analysis of fragments obtained by acid hydrolysis and Smith degradation of the O-polysaccharide, together with data on methylation analysis and nuclear magnetic resonance spectroscopic measurement of the polysaccharide, led to the most likely structure of the repeating units of the polymer chain, ----4)D-GalNAcA(alpha 1----3)D-QuiNAc(beta 1----2)L-Rha(alpha 1----3)L-Rha(alpha 1----, in which about 20% of the N-acetylgalactosaminuronic acid residues were in an amide form and about 75% of the same residues were O-acetylated at C-3.     

Structure of the lipopolysaccharide antigenic O-chain produced by Actinobacillus pleuropneumoniae serotype 4 (ATCC 33 378)

The structure of the antigenic O-polysaccharide part of the S-type lipopolysaccharide produced by Actinobacillus pleuropneumoniae serotype 4 has been determined by periodate oxidation, methylation, partial hydrolysis, and 1H- and 13C-n.m.r. spectroscopy. The O-polysaccharide structure has a branched-tetrasaccharide repeating unit, (----3)-beta-D-Galp-(1----4)-[beta-D-Glcp-(1----3)]-beta-D-GalpNAc- (1----4)-alpha-L-Rhap-(1-)n. The structure resembles that of the lipopolysaccharide O-chain of A. pleuropneumoniae serotype 7, and their common epitopes may account for the apparent serological cross-reactivity observed between the two serotypes when incompletely adsorbed, anticapsular-typing sera are used.     

The structure of the O-specific chain of lipopolysaccharide from Pseudomonas aeruginosa IID 1008 (ATCC 27584)

Structural studies were carried out on the O-polysaccharide fraction obtained from the lipopolysaccharide of Pseudomonas aeruginosa IID 1008 (ATCC 27584). The O-polysaccharide comprises L-rhamnose, N-acetyl-D-quinovosamine, N-acetyl-D-galactosaminuronic acid, and N-formyl-D-galactosaminuronic acid. The characterization of oligosaccharide fragments resulting from acid hydrolysis, Smith degradation and alkaline degradation of the O-polysaccharide, together with 1H-NMR and 13C-NMR spectroscopic data of the polysaccharide, led to the following structure for the repeating units: ----3)Rha(alpha 1----4)GalNAcA(alpha 1----4 GalNFoA(alpha 1----3)QuiNAc(alpha 1----. Almost all of the carboxyl groups of the N-acetylgalactosaminuronic acid residues and about half of the same groups of the N-formylgalactosaminuronic acid residues were in an amide form.     

Elucidation of the structure of the Pasteurella haemolytica serotype T10 lipopolysaccharide O-antigen by n.m.r. spectroscopy

The aqueous-phase lipopolysaccharide isolated from Pasteurella haemolytica serotype T10 cells by the phenol-water extraction method was found to be S-type lipopolysaccharide which possessed O-antigenic polysaccharide chains composed only of D-galactose residues. Structural analysis of the O-polysaccharide, using a combination of 1D and 2D 1H- and 13C-n.m.r. methods, led to the identification of the disaccharide repeating-unit as [----3)-alpha-D-Galp-(1----3)-beta-D-Galf-(1----]n. The serological cross-reactivity between P. haemolytica serotypes T4 and T10 can now be related to the structural similarity of the antigenic LPS O-polysaccharides.     

Novel blood group H glycolipid antigens exclusively expressed in blood group A and AB erythrocytes (type 3 chain H). I. Isolation and chemical characterization

A series of blood group H antigens reacting with monoclonal antibody MBrl has been found in human blood group A and AB erythrocytes, but not in O or B erythrocytes. These H antigens are clearly different from the globo-H structure (Fuc alpha 1----2Gal beta 1----3GalNAc beta 1----3Gal alpha 1----4Gal beta 1----4Glc beta 1----1Cer), which was previously isolated from O erythrocytes and is also reactive with the MBrl antibody. The new series of H antigens associated with blood group A has been characterized as having TLC mobilities which approximately coincide with those of H2, H3, and H4 glycolipids. One of these A-associated H antigens, having a similar TLC mobility as the H2 glycolipid, was isolated from A erythrocytes and was characterized by 1H NMR spectroscopy, methylation analysis, and enzymatic degradation as having the structure shown below: (formula, see text). The structure represents a precursor of the repetitive A epitope attached to type 2 chain, previously called type 3 chain A (Clausen, H., Levery, S. B., Nudelman, E., Tsuchiya, S., and Hakomori, S. (1985) Proc. Natl. Acad. Sci. U.S.A. 82, 1199-1203). This A-associated H structure is hereby called type 3 chain H.     

Serological diversity and chemical structures of Campylobacter jejuni low-molecular-weight lipopolysaccharides.

Low-Mr lipopolysaccharides (LPS) of Campylobacter jejuni reference strains for serotypes O:1, O:4, O:23, and O:36 were examined through the liberation of core oligosaccharides by mild acid cleavage of the ketosidic linkage of 3-deoxy-D-manno-2-octulosonic acid residues to the lipid A moiety. The liberated oligosaccharides were examined for chemical structure by compositional analysis and methylated linkage analysis in conjunction with fast atom bombardment-mass spectrometry of permethylated oligosaccharide derivatives. The results showed (i) that the LPS contained short oligosaccharide chains of branched nonrepetitive structure, to many of which N-acetylneuraminic acid residues remained attached by 2----3 linkages to 4-linked D-galactose residues in the core structure; (ii) that serotypical differences, which are not readily defined through qualitatively similar compositions, are clearly reflected in variations in linkage types and sequences of sugar residues in the outer core attached to an inner region of invariable structure; but (iii) that the presence or absence of NeuAc residues does not appear to be a basis for serotypical differences. The results also showed that oligosaccharide chains from LPS of serotypes O:1 and O:4 are distinctly different and are distinct again from those of the cross-reacting serotypes O:23 and O:36, between whose core oligosaccharide chains no differences were found. It is concluded that the structurally variable low-Mr LPS from C. jejuni show greater similarities to the lipooligosaccharides from Neisseria spp. than to the highly conserved core regions of Salmonella species. Those strains (serotypes O:23 and O:36) which also furnish high-Mr LPS are unique among gram-negative bacteria in possessing both low-Mr molecules of the Neisseria lipooligosaccharide type and high-Mr LPS of the Salmonella smooth type.     

Structural analysis of the O-antigen side chain polysaccharides in the lipopolysaccharides of Klebsiella serotypes O2(2a), O2(2a,2b), and O2(2a,2c).

The lipopolysaccharide (LPS) of Klebsiella serotype O2 is antigenically heterogeneous; some strains express multiple antigenic factors. To study this heterogeneity, we determined the structure of the O-antigen polysaccharides in isolates belonging to serotypes O2(2a), O2(2a,2b), and O2(2a,2c), by using composition analysis, methylation analysis, and both 1H and 13C nuclear magnetic resonance spectroscopy. The repeating unit structure of the 2a polysaccharide was identified as the disaccharide [----3)-beta-D-Galf-(1----3)-alpha-D-Galp-(1----] and was identical to D-galactan I, one of two O polysaccharides present in the LPS of Klebsiella pneumoniae serotype O1 (C. Whitfield, J. C. Richards, M. B. Perry, B. R. Clarke, and L. L. MacLean, J. Bacteriol. 173:1420-1431, 1991). LPS from serotype O2(2a,2b) also contained D-galactan I as the only O polysaccharide, suggesting that the 2b antigen is not an O antigen. The LPS of serotype O2(2a,2c) contained a mixture of two structurally distinct O polysaccharides and provides a second example of this phenomenon in Klebsiella spp. One polymer was identical to D-galactan I, and the other polysaccharide, the 2c antigen, was a polymer with a disaccharide repeating unit structure, [----3)-beta-D-GlcpNAc-(1----5)-beta-D-Galf-(1----]. The 2c structure does not resemble previously reported O polysaccharides from Klebsiella spp. Periodate oxidation confirmed that D-galactan I and the 2c polysaccharide are distinct glycans, rather than representing domains within a single polysaccharide chain. Monoclonal antibodies against the 2c antigen indicated that only LPS molecules with the longest O-polysaccharide chains contained the 2c epitope.     

Structure of the O-chain of the lipopolysaccharide of Pasteurella haemolytica serotype T3

Cell-wall lipopolysaccharide isolated from Pasteurella haemolytica serotype T3 using the phenol-water extraction procedure was shown to be an S type lipopolysaccharide by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Hydrolysis with mild acid afforded a lipid-free, antigenic O-chain polysaccharide. On the basis of one- and two-dimensional 1H and 13C nuclear magnetic resonance studies, in conjunction with microanalytical chemical methods, the O-polysaccharide was determined to be a linear polymer of a disaccharide repeating unit having the structure. [----3)-beta-D-G1cpNAc-(1----4)-alpha-L-Rhap-(1----]n     

Structure of the O-polysaccharide of Escherichia coli O112ab containing L-iduronic acid

An acidic O-polysaccharide was isolated by mild acid degradation of the lipopolysaccharide of Escherichia coli O112ab and studied by sugar analysis along with (1)H and (13)C NMR spectroscopy. The O-polysaccharide was found to contain a rarely occurring sugar component, L-iduronic acid (L-IdoA), and the following structure of the branched pentasaccharide repeating unit was established: [structure: see text].     

Structure of the group G streptococcal polysaccharide

The structure of the group-specific polysaccharide of group G Streptococcus was determined by means of methylation analysis and selective chemical degradations. The anomeric configurations and conformations of the sugar residues were studied by 1H- and 13C-n.m.r. spectroscopy. The tetrasaccharide repeating unit, ----3)-alpha-D-Galp-(1----2)-[alpha-L-Rhap-(1----3)-beta-D-GalpNAc - (1----4)]-alpha-L-Rhap-(1----, was determined.     

Structural characterization of a blood group A heptaglycosylceramide with globo-series structure. The major glycolipid based blood group A antigen of human kidney

A blood group A glycosphingolipid with the globo-series structure has been isolated from human kidney and structurally characterized. The structure was shown by mass spectrometry and proton NMR spectroscopy of the intact permethylated and permethylated-reduced derivatives together with degradation studies to be, GalNAc alpha 1----3Gal(2----1 alpha Fuc)beta 1----3GalNAc beta 1----3Gal alpha 1----4Gal beta 1----4Glc beta 1----1 Ceramide. This glycolipid reacts with both polyclonal and monoclonal anti-A blood group typing antisera and it is the major glycolipid based blood group A antigen present in the human kidney.     

Structural studies on the fucosamine-containing O-specific polysaccharide of Proteus vulgaris O19

The polysaccharide chain of Proteus vulgaris O19 lipopolysaccharide contains D-galactose, N-acetyl-D-glucosamine N-acetyl-D-galactosamine and N-acetyl-L-fucosamine in the ratio 1:1:1:1. The structure of the polysaccharide was established by full acid hydrolysis and methylation analysis, as well as by non-destructive methods, i.e. the computer-assisted evaluation of the 13C-NMR spectrum and computer-assisted evaluation of the specific optical rotation by Klyne's rule. The polysaccharide is regular and built up of tetrasaccharide repeating units of the following structure: ----3)-alpha-L-FucNAcp-(1----3)-beta-D-GlcNAcp-(1----3)-alph a-D-Galp- (1----4)-alpha-D-GalNAcp-(1---- The O19-antiserum cross-reacts with lipopolysaccharide from P. vulgaris O42, the structure of which is still unknown. No cross-reactions were observed with O-polysaccharides Pseudomonas aeruginosa O7 and Salmonella arizonae O59 in spite of some structural similarities.     

Structure and immunochemistry of an oligosaccharide repeating unit of the capsular polysaccharide of type III group B Streptococcus. A revised structure for the type III group B streptococcal polysaccharide antigen

We have derived oligosaccharides from the capsular polysaccharide of type III group B Streptococcus by enzymatic hydrolysis of a specific backbone glycosidic bond utilizing an endo-beta-galactosidase from Flavobacterium keratolyticus. Enzymatic digestion of the polysaccharide produced oligosaccharide fragments of one or more pentasaccharide repeating units. On the basis of 13C NMR, 1H NMR, and methylation analyses, it was established that the smallest digestion fragment was alpha-D-NeupNAc-(2----3)-beta-D-Galp-(1----4)-[beta-D-Glcp-(1----6 )]- beta-D-GlcpNAc-(1----3)-beta-D-Gal. The isolation of this oligosaccharide is consistent with the susceptibility of the beta-D-Galp-(1----4)-beta-D-Glcp linkage in the backbone of the type III group B streptococcal polysaccharide and confirms that the polysaccharide is composed of a pentasaccharide repeating unit. High resolution 13C NMR spectroscopic studies indicated that, as in the case of the pentasaccharide, the terminal sialic acid residues of the type III group B streptococcal polysaccharide were linked to O-3 and not to O-6 of its branch beta-D-galactopyranosyl residues as had been previously reported (Jennings, H. J., Rosell, K.-G., and Kasper, D. L. (1980) Can. J. Chem. 58, 112-120). This linkage was confirmed in an independent methylation analysis of the type III group B streptococcal polysaccharide. Thin layer chromatogram binding assay and radioactive antigen binding assays with radiolabeled oligosaccharides demonstrated the single repeating unit pentasaccharide oligosaccharide to be poorly antigenic. Increasing oligosaccharide size to a decasaccharide consisting of two repeating units resulted in an 8-fold increase in antigen binding in the direct radioactive antigen binding assay. The results suggest that a region of the immunodeterminant site critical for antibody binding is located in the backbone of the polysaccharide and involves the beta-D-galactopyranose-(1----4) beta-D-glucopyranose bond.