Somatic antigens of Pseudomonas aeruginosa. The structure of the O-specific polysaccharide chains of P. aeruginosa 011 (Lanyi) lipopolysaccharides

Abstract

Lipopolysaccharides were isolated from the phenol layer on aqueous phenol extraction of cells of Pseudomonas aeruginosa 011 (Lanyi classification), strains 170021 and 170040. On mild acid degradation of the lipopolysaccharides, with the subsequent gel-filtration on Sephadex G-50, neutral 0-specific polysaccharides made up of 6-deoxysugars alone were obtained. Two 2-acetamido-2,6-dideoxy-~-galactose (LFucNAc), 2-acetamido-2,6-dideoxy-D-glucose (DQuiNAc) and L-rhamnose (LRha) residues were found to be the components of the strain 170021 polysaccharide repeating units; those of strain 170040 contained the same monosaccharides, but, instead of 2-acetamido-2,6-dideoxy-~-glucose residue, that of 2-acetamido-2,6-dideoxy-~-galactose (DFucNAc) was present.On the basis of the I3C nuclear magnetic resonance data, methylation analysis and three successive Smith degradations the following structures were determined for the polysaccharide repeating units: strain 170021, +2)~Rha(al-+3)~FucNAc(ctl+3)~FucNAc(al+3)~QuiNAc(~1+ ; strain 170040, -+2)~Rha(al-t3)~FucNAc- More recent studies demonstrated a serological interrelationship between strain 170021 and serologically nonidentical to it strain 170040; in the new Lanyi and Bergan classification both of these strains were placed into one 0-serogroup and designated as serotypes 04a, b and 04a, c, respectively [l].In the present communication structures of the repeating units of the 0-specific polysaccharide of P. aeruginosa, strains 170021 and 170040, were established, and, on the basis of a marked structural similarity, the fact of their reference to the same 0-serogroup in the classification [l] was justified. MATERIALS AND METHODS Miscellaneous methodsInfrared spectra were measured with a UR-10 Karl Zeiss spectrometer in potassium bromide pellets. 'H and I3C nuclear magnetic resonance spectra were recorded with a WM- 250 (Bruker) instrument in D 2 0 at 60°C for polysaccharides and at 30 "C for oligosaccharides; chemical shifts are given from tetramethylsilane using as an internal standard acetone (6, = 2.23 ppm) or methanol (6, = 50.15 ppm).Optical rotations were determined with a Perkin-Elmer polarimeter, model 141, in water at 20°C. Solutions were freeze-dried or evaporated in vucuo at 40°C. Serological tests were conducted as described earlier [4]. Chromatography and electrophoresisAscending paper chromatography was carried out on Whatman paper in the system ethyl acetate/pyridine/water/ acetic acid (5/5/3/1, by vol.). Paper electrophoresis was performed in 0.025 M pyridine/acetate buffer, pH 4.5, at 28 V/cm. Alkaline silver nitrate reagent or ninhydrin solution were used to detect substances on paper.Gel-filtration was run on columns with Sephadex G-50 (67 x 3.3 cm or 38 x 1.3 cm) and Sephadex G-15 (87 x 1.4 cm). Gas-liquid chromatography was conducted on a Pye Unicam instrument series 104 (model 64), on a glass column (150x0.4cm) with 3% OV-1 on Diatomite CQ (100-200 mesh) ; carrier gas nitrogen. Gas-liquid chromatography/ mass spectrometry was performed on a Varian MAT 111...

Composition of an unusual accessory polymer containing N-lactyl-3-amino-3,6-dideoxyhexose from walls of Bacillus megaterium NCIB 7581

“…Ivatt & Gilvarg, 1978;Kaya et al, 1985). The linkage was also stable (shown by methods similar to those used for treatment with HF) to 4 M-NH,OH at 37 "C for 2 h. In contrast, Sasaki et al (1983) and Kojima et al (1985) successfully used similar conditions (0.5 M-NaOH at 37°C for 30min) to break the phosphodiester bonds between teichoic acids and peptidoglycan.…”

“…In certain actinomycetes, N-glycolyl-muramic acid is present (Adam et al, 1969). The lipopolysaccharide of some strains of Pseudomonas aeruginosa may contain N-formyl-2-amino-2-deoxygalacturonic acid (Knirel et al, 1985). Pseudaminic acid carries an N-3-hydroxybutyryl-substituent (Knirel et al, 1986).…”

“…The lipopolysaccharide of some strains of Pseudomonas aeruginosa may contain N-formyl-2-amino-2-deoxygalacturonic acid (Knirel et al, 1985). Pseudaminic acid carries an N-3-hydroxybutyryl-substituent (Knirel et al, 1986). An Nlactyl-aminosugar does not seem to have been described before the present work, although muramic acid lactam (Warth & Strominger, 1969) can be regarded as an Nlactyl-2-amino-2-deoxyhexose in which the lactyl-group is also linked to the aminosugar by an ether bond.…”

Outer Membrane Permeability of Pseudomonas aeruginosa

“…In recent years, further advances have been made in our understanding of the structure of the O-chain portion of this molecule by the work of Dmitriev, Kochetkov, and their co-workers (Dmitriev et al, 1980(Dmitriev et al, , 1982Knirel et al, 1982aKnirel et al, ,b, 1983. Even more important, these studies substantiated the earlier conclusions of Meadow and colleagues (Chester et al, 1973;Koval and Meadow, 1975) in showing that the serological classification of P. aeruginosa, Dmitriev et al (1980Dmitriev et al ( , 1982 and Knirel et al (1982aKnirel et al ( ,b, 1983. Even more important, these studies substantiated the earlier conclusions of Meadow and colleagues (Chester et al, 1973;Koval and Meadow, 1975) in showing that the serological classification of P. aeruginosa, Dmitriev et al (1980Dmitriev et al ( , 1982 and Knirel et al (1982aKnirel et al ( ,b, 1983.…”

Roles for wbtC, wbtI, and kdtA Genes in Lipopolysaccharide Biosynthesis, Protein Glycosylation, Virulence, and Immunogenicity in Francisella tularensis Strain SCHU S4

“…The OPS gene locus was annotated ( wbtA -to- wbtN ) and putative gene functions were assigned based upon homology to genes from other bacteria known to be involved in LPS biosynthesis [9]. In particular, many of the genes within the F. tularensis OPS locus were found to be homologous to the O -antigen genes of Pseudomonas aeruginosa serotype O6, which expresses a similar O -antigen repeat structure [22]. …”