Structure of the capsular polysaccharide of <i>Haemophilus pleuropneumoniae</i> serotype 5

Altman, Eleonora; Brisson, Jean‐Robert; Perry, Malcolm B.

doi:10.1111/j.1432-1033.1987.tb13685.x

Cited by 32 publications

(11 citation statements)

References 33 publications

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“…GlcN phosphate linked to Kdo at position 4, therefore, represents a novel type of substitution in lipopolysaccharides. The disaccharide bears some resemblance to the repeating units of the capsular antigens K14 and K15 of E. coli [12,131 and Actinobacillus (Haemophilus) pleuropneumoniae serotype 5a and 5b [14,151. These polymers, however, contain N-acetylated hexosamine, and the linkage is to position 5 of Kdo.…”

Section: Discussionmentioning

confidence: 99%

4‐O‐(2‐Amino‐2‐deoxy‐α‐d‐glucopyranosyl)‐3‐deoxy‐d‐manno‐2‐octulosonic acid, a constituent of lipopolysaccharides of the genus Pectinatus

Helander

Moll²,

Zähringer³

1993

European Journal of Biochemistry

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A disaccharide containing GlcN and 3-deoxy-D-manno-2-octu~osonic acid (Kdo) was detected after acidic methanolysis and peracetylation of hydrofluoric-acid-treated smooth-type and roughtype lipopolysaccharide of Pectinatus cerevisiiphilus and Pectinatus frisingensis, which are strictly anaerobic bacteria capable of growing in packaged beer. The disaccharide was also found in alkalitreated lipopolysaccharide, but was not directly detectable from intact lipopolysaccharide. This suggested that the disaccharide carried a phosphate residue. The position of this phosphate was shown, by GLCMS of appropriately degraded and derivatized samples, to be 0 6 of the GlcN. Methylation analysis of the purified disaccharide revealed that GlcN was linked to position 4 of Kdo. The acetylated derivative of the disaccharide was isolated in pure form, and, by 'H-NMR and I3C-NMR spectroscopy, it was confirmed to possess the structure a-~-GlcpN-(1'+4)-Kdo. In the lipopolysaccharide the amino group of GlcN is free.The Gram-negative bacterial genus Pectinatus contains two species, I? cerevisiiphilus and I? frisingensis. Both species are strict anaerobes, and they occur as contaminants and spoilage organisms in packaged beer [ 1, 21. The cell-surface lipopolysaccharides of the type strains of Pectinatus were recently reported to possess exceptional heterogeneity, and in fact two distinct lipopolysaccharide populations could be extracted from both species [3]. One of these lipopolysaccharides was of low-molecular-mass (rough, R) type, whereas the other contained additional sugar chains resembling smooth ( S ) lipopolysaccharide. Chemical characterization of the lipopolysaccharides indicated possible differences in the core oligosaccharide regions of the R and S type, particularly in the substitution of the 3-deoxyoctulosonic-acid region.These findings prompted us to structurally study the core region of Pectinatus lipopolysaccharides. In the present communication we describe a novel saccharide structure in the lipopolysaccharides of Pectinatus. This saccharide consists of a 3-deoxy-D-manno-2-octulosonic acid (Kdo) carrying at its position 4 an a-pyranosidic GlcN, which is substituted at position 6' by phosphate. MATERIALS AND METHODS Reference compoundsMethyl 2-acetarnido-3,4,6-triri-O-acetyl-2-deoxy-a-~-glucopyranoside was prepared from methyl 2-acetamido-2-de- oxy-a-D-glucopyranoside (a-D-GlcpNAc-OMe, Sigma) by standard derivatization (peracetylation) and purification (silica gel HPLC) procedures. Methyl (methyl 4,5,7,8-tetra-O-acetyl-3-deoxy-a-~-manno-2-octulopyranos~d)onate (aKdop-OMe methyl ester) was kindly provided by Prof. Paul Kosma, Universitat fur Bodenkultur, Vienna, Austria. Lipopoly saccharidesLipopolysaccharides were extracted from P. cerevisiiphilus ATCC 29359= and I? frisingensis ATCC 33332T by the phenol/chloroform/petroleum-ether method (5 : 5 :8, by vol. modification [4]) as described previously [3]. By sequential precipitations with water and acetone, R-type and S-type lipopolysaccharides, respectively, were obtained fr...

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Section: Discussionmentioning

confidence: 99%

4‐O‐(2‐Amino‐2‐deoxy‐α‐d‐glucopyranosyl)‐3‐deoxy‐d‐manno‐2‐octulosonic acid, a constituent of lipopolysaccharides of the genus Pectinatus

Helander

Moll²,

Zähringer³

1993

European Journal of Biochemistry

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Section: Methodsmentioning

confidence: 99%

“…The presence of DNA and proteins was analysed by electrophoresis and quantified using an ND-1000 spectrophotometer (NanoDrop). Total sugars were quantified using the method described by Altman et al (1987).Whole-genome sequencing and analysis. Sequencing of the complete genome of ibi-626 was done by BaseClear BV (Leiden, The Netherlands) using a Roche Illumina Genome Analyzer II.…”

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confidence: 99%

Selection of hyperadherent mutants in Pseudomonas putida biofilms

et al. 2011

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A number of genetic determinants required for bacterial colonization of solid surfaces and biofilm formation have been identified in different micro-organisms. There are fewer accounts of mutations that favour the transition to a sessile mode of life. Here we report the isolation of random transposon Pseudomonas putida KT2440 mutants showing increased biofilm formation, and the detailed characterization of one of them. This mutant exhibits a complex phenotype, including altered colony morphology, increased production of extracellular polymeric substances and enhanced swarming motility, along with the formation of denser and more complex biofilms than the parental strain. Sequence analysis revealed that the pleiotropic phenotype exhibited by the mutant resulted from the accumulation of two mutations: a transposon insertion, which disrupted a predicted outer membrane lipoprotein, and a point mutation in lapG, a gene involved in the turnover of the large adhesin LapA. The contribution of each alteration to the phenotype and the possibility that prolonged sessile growth results in the selection of hyperadherent mutants are discussed. INTRODUCTIONThe development of a multicellular community associated with a surface and surrounded by an exopolymeric matrix, referred to as a biofilm, is common to a variety of bacteria under different environmental conditions. Biofilm formation has received increasing attention due to its importance in medicine, since biofilm populations are considered relevant to chronic infection, and are more resistant to the action of antibiotics and biocidals than planktonic populations (Anderson & O'Toole, 2008;Høiby et al., 2010). Biofilm development is also relevant in industrial settings and in the design of bioreactors (Nicolella et al., 2000;Singh et al., 2006). Genetic determinants that play a role in biofilm formation have been unravelled in different bacterial species, and environmental and cellular signals that influence this process have also been described. These include iron availability, quorum sensing, and the intracellular secondary messenger cyclic di-GMP (Banin et al., 2006;Patriquin et al., 2008;Ueda & Wood, 2009;Coenye, 2010). Changes in the levels of cyclic di-GMP correlate with phenotypic changes associated with virulence, motility, colony morphology, production of exopolysaccharides, and the transition between planktonic and sessile lifestyles (Hengge, 2009; Römling & Simm, 2009, and references therein). Changes in the expression of different functions are also associated with one or the other lifestyle; in Pseudomonas aeruginosa and Pseudomonas putida, for example, differential expression of flagellar components has been observed when comparing planktonic and biofilm populations, and even during the various stages of biofilm development (Sauer & Camper, 2001;Sauer et al., 2002;Toutain et al., 2007). In the former organism, swarming motility and surface attachment are inversely regulated through a pathway that involves BifA, a cyclic-di-GMP phosphodiesterase, and the membra...

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“…It has been found in nature as part of the repeat units of some LPS O antigens in Providencia alcalifaciens and as the nonreducing chain terminating residue in the O12 antigen from Klebsiella pneumoniae (17,18). It has also been identified in CPS repeat units from E. coli serotype K12 (19), N. meningitidis group E (20), and Actinobacillus pleuropneumoniae 5a and 5b (21,22). However, the β-Kdo transferases required for synthesis of these structures have not been identified.…”

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confidence: 99%

KpsC and KpsS are retaining 3-deoxy- d - manno -oct-2-ulosonic acid (Kdo) transferases involved in synthesis of bacterial capsules

Willis

Whitfield

2013

Proc. Natl. Acad. Sci. U.S.A.

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Significance Capsules are bacterial surface structures used by many Gram-negative pathogens to evade the host immune system. They are comprised of long carbohydrate chains, called capsular polysaccharides (CPSs), which possess a lipid at one end. The lipid is connected to the CPS through an unusual linker consisting of five to nine residues of an acidic sugar 3-deoxy- d - manno -oct-2-ulosonic acid (Kdo). This study identifies two conserved proteins from CPS assembly systems, KpsC and KpsS, as the enzymes that synthesize the Kdo linker on the terminal lipid. Synthesis of this terminal glycolipid was reconstituted in vitro using purified enzymes and a synthetic lipid acceptor. The data support a unique model for CPS biosynthesis and identify these enzymes as unique targets for antibacterial therapies.

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Structure of the capsular polysaccharide of Haemophilus pleuropneumoniae serotype 5

Cited by 32 publications

References 33 publications

4‐O‐(2‐Amino‐2‐deoxy‐α‐d‐glucopyranosyl)‐3‐deoxy‐d‐manno‐2‐octulosonic acid, a constituent of lipopolysaccharides of the genus Pectinatus

4‐O‐(2‐Amino‐2‐deoxy‐α‐d‐glucopyranosyl)‐3‐deoxy‐d‐manno‐2‐octulosonic acid, a constituent of lipopolysaccharides of the genus Pectinatus

Selection of hyperadherent mutants in Pseudomonas putida biofilms

KpsC and KpsS are retaining 3-deoxy- d - manno -oct-2-ulosonic acid (Kdo) transferases involved in synthesis of bacterial capsules

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