Structure of the O-specific polysaccharide of the bacterium Proteus mirabilis O29.

An O-specific polysaccharide was obtained by mild acid degradation of P. mirabilis O29 lipopolysaccharide (LPS) and found to contain 2-acetamido-2-deoxy-D-galactose and D-glucuronic acid (D-GlcA) in the ratio 3:1. Studies of the polysaccharide by 1H- and 13C-NMR spectroscopy including two-dimensional correlation spectroscopy (COSY), total correlation spectroscopy (TOCSY), nuclear Overhauser effect spectroscopy (NOESY), and H-detected 1H,13C-heteronuclear multiple-quantum coherence (HMQC) experiments demonstrated the following structure of the branched tetrasaccharide repeating unit:

Structure of a glycerol teichoic acid-like O-specific polysaccharide of Proteus vulgaris O12.

A phosphorylated O-specific polysaccharide (O-antigen) was obtained by mild acid degradation of Proteus vulgaris O12 lipopolysaccharide and studied by sugar and methylation analyses, 1H-, 13C- and 31P-NMR spectroscopy, including two-dimensional COSY, TOCSY, NOESY, H-detected 1H, 13C and 1H, 31P heteronuclear multiple-quantum coherence experiments. It was found that the polysaccharide consists of pentasaccharide repeating units connected via a glycerol phosphate group, and has the following structure: where FucNAc is 2-acetamido-2,6-dideoxygalactose and the degree of O-acetylation at position 4 of GalNAc is approximately 25%. Immunochemical studies with P.

Structure of a 2-aminoethyl phosphate-containing O-specific polysaccharide of Proteus penneri 63 from a new serogroup O68.

Lipopolysaccharide of Proteus penneri strain 63 was degraded by mild acid to give a high molecular mass O-specific polysaccharide that was isolated by gel-permeation chromatography. Sugar and methylation analyses and NMR spectroscopic studies, including two-dimensional 1H, 1H COSY, TOCSY rotating-frame NOE spectroscopy, H-detected 1H,13C and 1H,31P heteronuclear multiple-quantum coherence (HMQC), and 1H, 13C HMQC-TOCSY experiments, demonstrated the following structure of the polysaccharide: where FucNAc is 2-acetamido-2,6-dideoxygalactose and PEtn is 2-aminoethyl phosphate. The polysaccharide studied shares some structural features, such as the presence of D-GlcNAc6PEtn and an alpha-L-FucNAc-(1-->3)-D-GlcNAc disaccharide, with other Proteus O-specific polysaccharides.

The O-chain polysaccharide of the lipopolysaccharide of Xanthomonas campestris pv. begoniae GSPB 525 is a partially L-xylosylated L-rhamnan.

The O-chain polysaccharide (OPS) of the lipopolysaccharide of Xanthomonas campestris pv. begoniae GSPB 525 was found to contain L-rhamnose and L-xylose in the ratio 1:0.6.

Chemical structure of a polysaccharide from Campylobacter jejuni 176.83 (serotype O:41) containing only furanose sugars.

A neutral polysaccharide was obtained by hot phenol-water extraction of biomass from Campylobacter jejuni 176.83 and subsequently separated from acid-liberated core oligosaccharide of lipopolysaccharide by sequential GPC on Bio-Gel P6 and TSK-40 columns. All sugar components of the trisaccharide repeating unit of the polysaccharide were found to be of the furanose ring form.

Structure and cross-reactivity of the O-antigen of Proteus vulgaris O8.

A high-molecular-mass O-specific polysaccharide was obtained by mild acid degradation of Proteus vulgaris O8 lipopolysaccharide followed by gel permeation chromatography. Studies of the polysaccharide by sugar and methylation analyses and 1H and 13C NMR spectroscopy, including 2D COSY, TOCSY, NOESY, and H-detected 1H, 13C heteronuclear multiple-quantum coherence (HMQC) experiments, demonstrated the presence of a tetrasaccharide repeating unit having the following structure: [sequence: see text] The role of an epitope associated with the alpha-L-FucpNAc-(1-->3)-D-GlcpNAc disaccharide in serological cross-reactivity of P. vulgaris O8 is discussed.

Structure of a new acidic O-antigen of Proteus vulgaris O22 containing O-acetylated 3-acetamido-3,6-dideoxy-D-glucose.

The acidic O-specific polysaccharide of Proteus vulgaris O22 was studied using 1H and 13C NMR spectroscopy, including 2D COSY, TOCSY, NOESY, and H-detected 1H, 13C heteronuclear multiple-quantum coherence (HMQC) experiments, and the following structure for the branched pentasaccharide repeating unit was established: [sequence: see text] where Qui3NAc is 3-acetamido-3,6-dideoxyglucose, O-acetylation of QuiNAc at position 4 is stoichiometric and at position 2 nonstoichiometric. Serological relationships of P. vulgaris O22 with some other Proteus strains were substantiated on the level of the O-antigen structures.

Structures of the O-specific polysaccharides and a serological cross-reactivity of the lipopolysaccharides of Proteus mirabilis O24 and O29.

Strains of Proteus mirabilis belonging to serogroups O24 and O29 are frequent in clinical specimens. Anti-P. mirabilis O24 serum cross-reacted with the lipopolysaccharide (LPS) of P.

Structural and serological studies on the O-antigen of Proteus mirabilis O14, a new polysaccharide containing 2-[(R)-1-carboxyethylamino]ethyl phosphate.

An O-specific polysaccharide was obtained by mild acid degradation of Proteus mirabilis O14 lipopolysaccharide (LPS) and found to contain D-galactose, 2-acetamido-2-deoxy-D-glalactose, phosphate, N-(2-hydroxyethyl)-D-alanine (D-AlaEtn), and O-acetyl groups. Studies of the initial and O-deacetylated polysaccharides using one- and two-dimensional 1H- and 13C-NMR spectroscopy, including COSY, TOCSY, NOESY, H-detected 1H,13C heteronuclear multiple-quantum coherence, and heteronuclear multiple-bond correlation experiments, demonstrated the following structure of the repeating unit: [equation: see text] This is the second bacterial polysaccharide reported to contain alpha-D-Galp6PAlaEtn, whereas the first one was the O-antigen of P. mirabilis EU313 taken erroneously as strain PrK 6/57 from the O3 serogroup [Vinogradov, E.

Serological studies of an acid-labile O-polysaccharide of Proteus vulgaris OX19 lipopolysaccharide using human and rabbit antibodies.

In a Weil-Felix test, sera from patients infected with Rickettsia sp. agglutinate Proteus OX types of bacteria and Proteus lipopolysaccharide (LPS) are responsible for the cross-reaction. Data on the character of LPS of one of the OX group strains, Proteus vulgaris OX19, are contradictory, and it remained unclear whether it has an O-polysaccharide (OPS) and is thus LPS of the smooth type (S) or not (rough-type LPS).

More on the structure of Vibrio cholerae O22 lipopolysaccharide.

The structure of a short-chain lipopolysaccharide (LPS) of Vibrio cholerae O22 strain 169-68, that cross-reacts with V. cholerae O139 Bengal, was elucidated. The structure differs in detail from that reported on another strain of O22 [A.

Structure of the acidic polysaccharide chain of the lipopolysaccharide of Shewanella alga 48055.

A lipopolysaccharide (LPS) with an acidic polysaccharide chain was isolated from the bacterium Shewanella alga strain 48055 and cleaved selectively at the glycosidic linkage of N-acetylneuraminic acid to give a tetrasaccharide. Studies of the tetrasaccharide and the O-deacylated LPS by 1H and 13C NMR spectroscopy, including 2D COSY, TOCSY, NOESY, rotating-frame NOE spectroscopy (ROESY), and H-detected 1H, 13C heteronuclear multiple-quantum coherence (HMQC) experiments, revealed the following structure of the polysaccharide repeating unit: -->3)-beta-D-GalpA6GroN-(1-->3)-beta-D-GlcpNAc-(1-->3)-alpha-D- GalpA6GroN- (1-->4)-alpha-Neup5Ac-(2--> where GroN is an amidically linked residue of 2-amino-1,3-propanediol (2-amino-2-deoxyglycerol). A similar structure, but with 2-acetamido-2,6-dideoxy-D-glucose instead of 2-acetamido-2-deoxy-D-glucose, has been reported previously for the polysaccharide chain of a non-O1 Vibrio cholerae H11 LPS [E.

Structure of a highly acidic O-specific polysaccharide of lipopolysaccharide of Pseudoalteromonas haloplanktis KMM 223 (44-1) containing L-iduronic acid and D-QuiNHb4NHb.

An acidic O-specific polysaccharide was obtained by mild acid degradation of the lipopolysaccharide isolated by phenol-water extraction of Pseudoalteromonas haloplanktis strain KMM 223 (44-1). L-Iduronic acid (IdoA) was found to be a component of the polysaccharide and identified by NMR spectroscopy and after carboxyl-reduction followed by acid hydrolysis and acetylation, by GLC-MS as 2,3,4-tri-O-acetyl-1,6-anhydroidose. On the basis of 1H and 13C NMR spectroscopic studies, including 1D NOE, 2D NOESY, HSQC and HMBC experiments, the following structure of the branched pentasaccharide repeating unit of the polysaccharide was established: -->4)-beta-D-GlcpAI-(1-->4)-beta-D-GlcpAII-(1-->3)-beta-D-++ +QuipNHb4NHbII- (1-->2)-alpha-L-IdopA-(-->4 increases 1 alpha-D-QuipNAc4NAcI where QuiNAc4NAc and QuiNHb4NHb are 2,4-diacetamido-2,4,6-trideoxyglucose and 2,4,6-tri-deoxy-2,4- di[(S)-3-hydroxybutyramido]glucose, respectively.

Structure of the O-specific polysaccharide of Proteus penneri strain 41 from a new proposed serogroup O62.

O-specific polysaccharide of Proteus penneri strain 41 was studied using 1H- and 13C-NMR spectroscopy, including two-dimensional COSY, heteronuclear 13C,1H-correlation (HETCOR) and one-dimensional NOE spectroscopy, and the following structure of a non-stoichiometrically O-acetylated hexasaccharide repeating unit was established:[structure: see text] where RGlcNAc is 2-acetamido-4-O-[(S)-1-carboxyethyl]-2-deoxyglucose. Cross-reactivity of anti-P. penneri 41 O-serum with other P.

Structure of the O-antigen of Vibrio cholerae O155 that shares a putative D-galactose 4,6-cyclophosphate-associated epitope with V. cholerae O139 Bengal.

The O-specific polysaccharide of Vibrio cholerae 0155 was studied by sugar and methylation analyses, dephosphorylation with 48% hydrofluoric acid, 1H- and 13C-NMR spectroscopy, including two-dimensional COSY, TOCSY, NOESY, and heteronuclear single-quantum coherence (HSQC) experiments. The following structure of the pentasaccharide repeating unit of the polysaccharide was established: carbohydrate sequence [see text]. An unusual component, D-galactose 4,6-cyclophosphate, has been reported previously as a component of the capsular polysaccharide and O-antigen of V.

Structure of a neutral O-specific polysaccharide of the bacterium Proteus mirabilis O24.

Lipopolysaccharide of the bacterium Proteus mirabilis O24 was found to have a neutral O-specific polysaccharide chain containing D-galactose, 2-acetamido-2-deoxy-D-glucose, and 2-acetamido-2-deoxy-D-galactose in ratios 1:2:1. On the basis of 1H- and 13C-NMR spectroscopy, including two-dimensional correlation spectroscopy (COSY), H-detected 1H, 13C heteronuclear multiple-quantum coherence (HMQC), and nuclear Overhauser effect spectroscopy (NOESY), the following structure of the branched tetrasaccharide repeating unit of the O-specific polysaccharide was established: -->3)-beta-D-GlcpNAc-(1-->4)-beta-D-GalpNAc-(1-->4)-beta-D-GlcpNAc-(1--> [formula: see text] beta-D-Galp.

Structures of the O-antigens of Proteus bacilli belonging to OX group (serogroups O1-O3) used in Weil-Felix test.

Structures of the O-specific polysaccharide chains of lipopolysaccharides from Proteus group OX strains (serogroups O1-O3) used as antigens in Weil-Felix test for diagnosis of rickettsiosis, were established. From them, the acid-labile polysaccharide of Proteus vulgaris OX19 (O1) is built up of the following branched pentasaccharide repeating units connected via a phosphate group: [structure in text] where QuiNAc stands for 2-acetamido-2,6-dideoxyglucose (N-acetylquinovosamine). The basis of serospecificity of the Proteus group OX antigens and their cross-reactivity with human anti-rickettsial antibodies is discussed.

Structural studies on the short-chain lipopolysaccharide of Vibrio cholerae O139 Bengal.

A Vibrio cholerae O139 strain, MO10-T4, lacking capsular polysaccharide, produces a short-chain lipopolysaccharide (LPS), similar to enterobacterial SR strains. It was studied by acidic and alkaline degradation, dephosphorylation, sugar and methylation analysis, high-performance anion-exchange chromatography, one- and two-dimensional 1H-, 13C-, and 31P-NMR spectroscopy, and electrospray ionization mass spectrometry. The following structure was proposed for the core region of the LPS: [structure: see text] where PEtn stands for 2-aminoethyl phosphate, Fru for fructose, Hep for L-glycero-D-manno-heptose, and Kdo for 3-deoxy-D-manno-octulosonic acid; unless otherwise stated, the monosaccharide residues are D and present in the pyranose form.

Structure of the O-specific polysaccharide of Salmonella enterica ssp. arizonae O50 (Arizona 9a,9b).

On the basis of sugar and methylation analysis, selective removal of 3,6-dideoxy-L-xylohexose (colitose, Col), 1H and 13C NMR spectroscopy, including 1D NOE, 2D COSY, and 2D H-detected 1H, 13C heteronuclear multiple-quantum coherence (HMQC), the following structure of the repeating unit of the O-specific polysaccharide of Salmonella enterica ssp. arizonae O50 (Arizona 9a,9b) was established: [sequence: see text] The O-antigen studied includes a trisaccharide fragment alpha-Co1p-(1-->2)-beta-D-Galp-(1-->3)-beta-D-GlcpNAc, which is a colitose ('3-deoxy-L-fucose') analogue of the Lewis (precursor) blood group antigen.

Structure of a new 6-deoxy-alpha-D-talan from Burkholderia (Pseudomonas) plantarii strain DSM 6535, which is different from the O-chain of the lipopolysaccharide.

An O-acetylated homopolysaccharide of 6-deoxy-D-talose (6-deoxy-alpha-D-talan polymer) was isolated from Burkholderia (Pseudomonas) plantarii strain DSM 6535 by extraction with 2-propanol. The structure (1) of the trisaccharide repeating unit of the polysaccharide was established by studies of the intact and O-deacetylated polysaccharides using methanolysis, methylation analysis, 1H and 13C NMR spectroscopy, including 2D COSY, heteronuclear 13C, 1H COSY, 1D NOE, and computer-assisted analysis of 1D 13C NMR spectra. The remaining material after extraction of the biomass with 2-propanol showed to be a lipopolysaccharide with an O-specific polysaccharide chain having a different structure (2), which has been found previously in lipopolysaccharides of a number of other Gram-negative bacteria.

Structure of the O-specific polysaccharide of the bacterium Proteus mirabilis O13 containing a novel component: an amide of D-galacturonic acid with N(epsilon)-(1-carboxyethyl)lysine.

An acidic O-specific polysaccharide was obtained by mild degradation of the lipopolysaccharide of the bacterium Proteus mirabilis O13 and found to contain D-galactose, 2-acetamido-2-deoxy-D-glucose, and N(epsilon)-(1-carboxyethyl)-N(alpha)-(D-galacturonoyl)lysine. On the basis of full acid hydrolysis and 1H- and 13C-NMR spectroscopy, including two-dimensional correlation spectroscopy (COSY), H-detected heteronuclear 1H, 13C multi-quantum coherence (HMQC), and rotating-frame nuclear Overhauser effect spectroscopy (ROESY), the following structure of the branched trisaccharide repeating unit of the polysaccharide was established [structure: see text]

Structure of the acid-labile galactosyl phosphate-containing O-antigen of the bacterium Proteus vulgaris OX19 (serogroup O1) used in the Weil-Felix test.

The structure of the O-specific polysaccharide chain of Proteus vulgaris OX19 lipopolysaccharide which determines the O1 specificity of Proteus and is used in the Weil-Felix test for diagnostics of rickettsiosis was established. On the basis of 1H- and 13 C-NMR spectroscopy, including two-dimensional correlation spectroscopy (COSY), H-detected 1H, 13C heteronuclear multiple-quantum coherence (HMQC), and rotating-frame nuclear Overhauser effect spectroscopy (ROESY), it was found that the polysaccharide consists of branched pentasaccharide repeating units containing D-galactose, 2-acetamido-2-deoxy-D-glucose, 2-acetamido-2-deoxy-D-galactose, and 2-acetamido-2,6-dideoxy-D-glucose (QuiNAc, two residues), which are connected to each other via a phosphate group (P): [formula: see text]. The polysaccharide is acid-labile, the glycosyl phosphate linkage being cleaved at pH 4.

The O-specific polysaccharide chain of Campylobacter fetus serotype A lipopolysaccharide is a partially O-acetylated 1,3-linked alpha-D-mannan.

A polysaccharide fraction liberated from Campylobacter fetus subsp. fetus serotype A lipopolysaccharide by mild acid hydrolysis followed by gel-permeation chromatography contained a partially O-acetylated D-mannan chain, as an O-specific polysaccharide, with a core oligosaccharide attached. The structure of the polysaccharide was studied by O-deacetylation, methylation, and 1H- and 13C-NMR spectroscopy, including computer-assisted analysis of the 13C-NMR spectrum.

Structural and serological studies of the O-antigen of the bacterium Proteus mirabilis OXK (serogroup O3) used in the Weil-Felix test.

Sugar analysis and 1H- and 13C-NMR spectroscopic studies showed that various strains of Proteus mirabilis OXK used as antigens in the Weil-Felix test for serodiagnosis of rickettsiosis (scrub typhus) produce lipopolysaccharides (LPSs) with the same O-specific polysaccharide chain having the following structure: [formula: see text] where GlcA and GalA are glucuronic and galacturonic acids, respectively. This polysaccharide which defines the O3 specificity of Proteus and has been found earlier in an unclassified P. mirabilis strain S1959, contains an amide of D-galacturonic acid with L-lysine which plays an important role in manifesting the immunospecificity.

Structural and serological studies of the O-antigen of the bacterium Proteus vulgaris OX2 (serogroup O2) used in the Weil-Felix test.

Based on monosaccharide analysis and 1H- and 13C-NMR spectroscopy, the following structure of the O-specific polysaccharide chain of Proteus vulgaris OX2 lipopolysaccharide (LPS), which defines the O2 specificity of Proteus, was established: [formula: see text] where L-QuiNAc is N-acetyl-L-quinovosamine (2-acetamido-2,6-dideoxy-L-glucose). Various strains of P. vulgaris OX2 used in the Weil-Felix test for serodiagnosis of rickettsiosis (spotted fevers, except for Rocky Mountain spotted fever) were shown to produce LPS with the same O-specific polysaccharide, which differs structurally and serologically from LPS of P.