Somatic antigens of Pseudomonas aeruginosa. The structure of the O-specific polysaccharide chains of lipopolysaccharides of P. aeruginosa serogroup O4 (Lanyi) and related serotype O6 (Habs) and immunotype 1 (Fisher)

Knirel, Yuriy A.; Vinogradov, Evgeny; Shashkov, Alexander S.; Dmitriev, Boris A.; Kochetkov, Nikolay K.; Stanislavsky, Evgeny S.; Mashilova, Galina M.

doi:10.1111/j.1432-1033.1985.tb09055.x

Cited by 48 publications

(35 citation statements)

References 33 publications

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“…The position of the signals for C-5 and C-7 in the region of 46 -54 ppm showed that these carbons carried nitrogen, and the chemical shifts of the signals for C-3 (34.8 ppm) and C-9 (16.2-16.3 ppm) indicated that these carbons entered the deoxy units. The position of the signals of xylose in the spectra of [1][2][3] were in good agreement with the data for methyl B-D-xylopyranoside [24], and hence this monosaccharide occupied the non-reducing terminus of the oligosaccharides. The use of the 13C nuclear magnetic resonance data for the determination of the mode of glycosidation and substitution by 0-acyl and N-acyl groups in I -3 was described above.…”

Section: Interpretation Of 3c Nuclear Magnetic Resonance Spectrasupporting

confidence: 86%

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Somatic antigens of Pseudomonas aeruginosa. The structure of O-specific polysaccharide chains of the lipopolysaccharides from P. aeruginosa O5 (Lanyi) and immunotype 6 (Fisher)

et al. 1987

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Lipolysaccharides were isolated from dry bacterial cells of Pseudomonas aeruginosa OSa,b,c, OSa,b,d, 05a,d (Lanyi classification) and immunotype 6 (Fisher classification) by the Westphal procedure. Their polysaccharide chains were built up of trisaccharide repeating units containing ~-xylose, 2-acetamido-2,6-dideoxy-~-galactose and a new sialic acid-like sugar, the di-N-acyl derivative of 5,7-diarnino-3,5,7,9-tetradeoxy-~-glycero-~-mannononulosonic (pseudaminic) acid. Formyl, acetyl and (R)-3-hydroxybutyryl groups were identified as the N-acyl substituents of the last monosaccharide; O5a,b,c and O5a,b,d lipopolysaccharides also contained 0-acetyl groups.The glycosidic linkage of pseudaminic acid was extremely labile towards acids, and mild acid degradation of the lipopolysaccharides produced, instead of the 0-specific polysaccharides, their trisaccharide fragments with pseudaminic acid at the reducing terminus. Similar degradation of immunotype 6 lipopolysaccharides, followed by oxidation with sodium metaperiodate, resulted in a disaccharide fragment due to destruction of xylose. In contrast the glycosidic linkage of pseudaminic acid proved to be more stable towards treatment with hydrogen fluoride than those of xylose and N-acetylfucosamine. As a result, solvolysis of immunotype 6 lipopolysaccharide with hydrogen fluoride in methanol gave methyl glycosides of a disaccharide and a trisaccharide with pseudaminic acid at the non-reducing terminus. Mild acid hydrolysis of these oligosides afforded free 5-N-acetyl-7-Nformylpseudaminic acid, which was identified by the 'H and 13C nuclear magnetic resonance data, as well as by the mass spectrum of the corresponding fully methylated aldonic acid.As a result of the identification of all oligosaccharides obtained and comparative analysis of the 13C nuclear magnetic resonance spectra of the oligosaccharides and lipopolysaccharides the following structures were estab- +~) -D -X~~~-(~~+~) -D -F~C N A C -(~~+~) -P S~~N A C~N F~-(~where D-xyl = ~-xylose, D-FUCNAC = 2-acetamido-2,6-dideoxy-~-galactose, Pse5N7NFm = 5-amino-3,5,7,9-tetradeoxy-7-formamido-~-glycero-~-manno-nonulosonic acid (7-N-formylpseudaminic acid). All the polysaccharides have an identical carbohydrate skeleton and differ from each other by the acyl substituent at N-5 of pseudaminic acid [acetyl or (R)-3-hydroxybutyryl group] or by the presence or absence of the 0-acetyl group at position 4 of N-acetylfucosamine. The data obtained account properly for the 0 specificity of the studied P. aeruginosa strains.The data on composition and structure of lipopolysaccharides, determining the 0 specificity of gram-negative bacteria, are employed to work out intraspecies chemotype schemes. Investigation of the lipopolysaccharides of an opportunistic bacterial pathogen P. aeruginosa showed many [2, 81, acetamidino [5, 6, 91, (R)- [7] and (S)-3-hydroxybutyryl [3] groups.In the present communication we report the establishing of the structures of the 0-specific polysaccharide chains of P. aeruginosa 0 5 (Lanyi classifi...

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Section: Interpretation Of 3c Nuclear Magnetic Resonance Spectrasupporting

confidence: 86%

“…Chemical shift (pprn) 170 Attempts to remove completely the 0-acetyl group by treatment of OSa,b,d lipopolysaccharide with aqueous triethylamine [2] failed. The 3C nuclear magnetic resonance data showed that even under rather drastic conditions (lOOT, 4 h) the degree of the-0-acetylation was not greater than 60%.…”

Section: _mentioning

confidence: 99%

Somatic antigens of Pseudomonas aeruginosa. The structure of O-specific polysaccharide chains of the lipopolysaccharides from P. aeruginosa O5 (Lanyi) and immunotype 6 (Fisher)

et al. 1987

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“…It is known that in a1 -3-linked disaccharides with the glycosidated pyranose having the D-ghC0 configuration, the effect on C-4 of this pyranose is of small negative or positive value and does not exceed 1 ppm by module if the glycosidating pyranose has the D configuration. In contrast, in the case of the L configuration of the glycosidating pyranose the effect on C-4 is negative and greater than 1 ppm by module [22]. For disaccharide 1 this effect amounts to +0.7 to +0.9 ppm (as was found by comparison with the I3C-NMR data for N-acetyl-Dquinovosamine [20]), and hence N-acetylgalactosaminuronic acid has the D configuration.…”

Section: Solvolysis With Hydrogen Fluoride; Oligosaccharidementioning

confidence: 77%

“…Their acidic properties are stipulated by the derivatives of amino sugars belonging to three classes: 2-amino-2-deoxyuronic acids [22,23,30, 311, 2,3-diamino-2,3-dideoxyuronic acids [15, 321, and 5,7-diamino-3,5,7,9-tetradeoxynonulosonic acids [20,33,34]. The polysaccharides of P. aeruginosa I1 (Sandvik) and V (VerderEvans) proved to be acidic as well owing to the presence of N-acetyb-galactosaminuronic acid.…”

Section: Discussionmentioning

confidence: 99%

“…The polysaccharides of P. aeruginosa I1 (Sandvik) and V (VerderEvans) proved to be acidic as well owing to the presence of N-acetyb-galactosaminuronic acid. Among the derivatives of this class N-acetyl-L-galactosaminuronic acid [23,301, Nformyl-D-galactosaminuronic acid [22], N-acetyl and Nformyl-D-galactosaminuronamides [22,311 were identified formerly in different P . aeruginosa 0-specific polysaccharides.…”

Section: Discussionmentioning

confidence: 99%

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Somatic antigens of Pseudomonas aeruginosa. The structure of O-specific polysaccharide chains of the lipopolysaccharides from P. aeruginosa II (Sandvik) and V (IM-1, Verder-Evans)

et al. 1987

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The lipopolysaccharides from two serologically related strains of Pseudomonas aeruginosa I1 (Sandvik classification) and V (IM-1, Verder-Evans classification) were isolated by the Westphal method and cleaved with 1 % acetic acid. The 0-specific polysaccharide from Sandvik I1 lipopolysaccharide involved L-rhamnose, N-acetyl-D-quinovosamine, and N-acetyl-D-galactosaminuronic acid in the ratio 2: 1 : 1, as well as 0-acetyl groups. The Verder-Evans V 0-specific polysaccharide was found to contain just the same sugars and, in addition, D-glucose, the monosaccharide ratio being 2: 1 : 1 : 1. On the basis of methylation analysis, Smith degradation, solvolysis with anhydrous hydrogen fluoride and 'H and I3C nuclear magnetic resonance data, it was concluded that the polysaccharides have identical main chains, and there were established the following structures of their repeating units:The data obtained allow us to substantiate the combining of the strains studied into one 0-serogroup as two individual 0-serotypes.An opportunistic pathogen Pseudomonas aeruginosa is known as a serologically heterogeneous species, and many serological schemes have been proposed for its classification (reviews [l, 21). On the basis of the classifications of Linyi [3] and Habs [4] the new most complete scheme has been compiled 111. In establishing the correlation between the strains from various classifications [I] it was found that the serogroup I1 of Sandvik's scheme [5] and the serogroup V (IM-1) of Verder-Evans' scheme [6], in contrast to the other groups of schemes, lack any counterparts in the classifications of Lanyi and Habs. Also, the serogroups SandvikII and Verder-Evans V turned out to be serologically interrelated, and were combined in the new compiled scheme into one serogroup 013 as serotypes 013a,13b and 013a,13c respectively.Immunospecificity of gram-negative bacteria is defined by the structure of polysaccharide chain of 0-antigens and, thus, chemical and immunochemical studies of P. aeruginosa lipopolysaccharides should provide a molecular basis for the classification of strains of this species. The present work Correspondence to Yu. A. Knirel, Institut Organicheskoj Khimii imeni N. D. Zelinskogo, Akademiya Nauk S.S.S.R., Leninskjj prospekt 47, Moskva, USSR-117913Abbreviations. QuiNAc, 2-acetamido-2,6-dideoxyglucose (Nacetylquinovosamine); GalNAcA, 2-acetamido-2-deoxygalacturonic acid (N-acetylgalactosaminuronic acid). deals with the establishing of the structures of 0-specific polysaccharides of P. aeruginosa I1 (Sandvik) and V (VerderEvans). MATERIALS AND METHODS Miscellaneous methods'H and I3C nuclear magnetic resonance (NMR) spectra were run on WM-250 and AM-300 (Bruker) instruments, respectively, with samples in D 2 0 at 60 "C (polysaccharides) and 30°C (oligosaccharides) and acetone (6, = 2.23 ppm) or methanol (6, = 50.15 ppm) as internal standard. Optical rotations were measured with a polarimeter EPO-1 (USSR) and solutions in water at 20°C. Solutions were freeze-dried or rotary evaporated in vacuo at 40°C. Methy...

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Three‐dimensional structure of a sugar N‐formyltransferase from Francisella tularensis

2014

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N-formylated sugars have been observed on the O-antigens of such pathogenic Gram-negative bacteria as Campylobacter jejuni and Francisella tularensis. Until recently, however, little was known regarding the overall molecular architectures of the N-formyltransferases that are required for the biosynthesis of these unusual sugars. Here we demonstrate that the protein encoded by the wbtj gene from F. tularensis is an N-formyltransferase that functions on dTDP-4-amino-4,6-dideoxy-D-glucose as its substrate. The enzyme, hereafter referred to as WbtJ, demonstrates a strict requirement for N 10 -formyltetrahydrofolate as its carbon source. In addition to the kinetic analysis, the three-dimensional structure of the enzyme was solved in the presence of dTDP-sugar ligands to a nominal resolution of 2.1 Å . Each subunit of the dimeric enzyme is dominated by a "core" domain defined by Met 1 to Ser 185. This core motif harbors the active site residues. Following the core domain, the last 56 residues fold into two a-helices and a b-hairpin motif. The hairpin motif is responsible primarily for the subunit:subunit interface, which is characterized by a rather hydrophobic pocket. From the study presented here, it is now known that WbtJ functions on C-4 0 amino sugars. Another enzyme recently investigated in the laboratory, WlaRD, formylates only C-3 0 amino sugars. Strikingly, the quaternary structures of WbtJ and WlaRD are remarkably different. In addition, there are several significant variations in the side chains that line their active site pockets, which may be important for substrate specificity. Details concerning the kinetic and structural properties of WbtJ are presented.

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Somatic antigens of Pseudomonas aeruginosa. The structure of the O-specific polysaccharide chains of lipopolysaccharides of P. aeruginosa serogroup O4 (Lanyi) and related serotype O6 (Habs) and immunotype 1 (Fisher)

Abstract: The structure of the 0-specific polysaccharide chains of lipopolysaccharides of P. aeruginosa serogroup 0 4 (Lanyi) and related serotype 0 6 (Habs) and immunotype 1 (Fisher)

Cited by 48 publications

References 33 publications

Somatic antigens of Pseudomonas aeruginosa. The structure of O-specific polysaccharide chains of the lipopolysaccharides from P. aeruginosa O5 (Lanyi) and immunotype 6 (Fisher)

Somatic antigens of Pseudomonas aeruginosa. The structure of O-specific polysaccharide chains of the lipopolysaccharides from P. aeruginosa O5 (Lanyi) and immunotype 6 (Fisher)

Somatic antigens of Pseudomonas aeruginosa. The structure of O-specific polysaccharide chains of the lipopolysaccharides from P. aeruginosa II (Sandvik) and V (IM-1, Verder-Evans)

Three‐dimensional structure of a sugar N‐formyltransferase from Francisella tularensis

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