Structures of the core oligosaccharide and O-units in the R- and SR-type lipopolysaccharides of reference strains of<i>Pseudomonas aeruginosa</i>O-serogroups

Bystrova, Olga V.; Knirel, Yuriy A.; Lindner, Buko; Kocharova, Nina A.; Kondakova, Anna N.; Zähringer, Ulrich; Pier, Gerald B.

doi:10.1111/j.1574-695x.2005.00004.x

Cited by 60 publications

(94 citation statements)

References 33 publications

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“…As expected, the rmlC mutant core OS structure lacked L-Rha residues. The presence of partial acetylation (50 to 70%) of the outer core sugars, such as residue L, found in LPS from rmlC, wapR, and migA rmd knockout strains and the phosphorylation patterns of all strains were consistent with the results described in a previous report (2).…”

Section: Discussionsupporting

confidence: 80%

“…The outer core is composed of four D-glucose (D-Glc) residues, one L-rhamnose (L-Rha) residue, and one N-(L-alanyl)-D-galactosamine residue, and its biosynthesis is not fully understood. The structures of the core OS of several different serotypes, clinical isolates, and rough mutants of P. aeruginosa have been reported previously (2,3,12,17,24). The results of the previous studies showed that there are minor variations in phosphate and O-acetyl substituents in some of the core sugars.…”

mentioning

confidence: 67%

“…All enzymes were used according to the manufacturers' instructions. Plasmid DNA was introduced into E. coli and P. aeruginosa by transformation (2). All constructs were verified by nucleotide sequencing at the Laboratory Services Division of the University of Guelph (Guelph, Ontario, Canada).…”

Section: Methodsmentioning

confidence: 99%

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Functional Characterization of MigA and WapR: Putative Rhamnosyltransferases Involved in Outer Core Oligosaccharide Biosynthesis ofPseudomonas aeruginosa

et al. 2008

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Pseudomonas aeruginosa lipopolysaccharide (LPS) contains two glycoforms of core oligosaccharide (OS); one form is capped with O antigen through an ␣-1,3-linked L-rhamnose (L-Rha), while the other is uncapped and contains an ␣-1,6-linked L-Rha. Two genes in strain PAO1, wapR (PA5000) and migA (PA0705), encode putative glycosyltransferases associated with core biosynthesis. We propose that WapR and MigA are the rhamnosyltransferases responsible for the two linkages of L-Rha to the core. Knockout mutants with mutations in both genes were generated. The wapR mutant produced LPS lacking O antigen, and addition of wapR in trans complemented this defect. The migA mutant produced LPS with a truncated outer core and showed no reactivity to outer core-specific monoclonal antibody (MAb) 5C101. Complementation of this mutant with migA restored reactivity of the LPS to MAb 5C101. Interestingly, LPS from the complemented migA strain was not reactive to MAb 18-19 (specific for the core-plus-one O repeat). This was due to overexpression of MigA in the complemented strain that caused an increase in the proportion of the uncapped core OS, thereby decreasing the amount of the core-plus-one O repeat, indicating that MigA has a regulatory role. The structures of LPS from both mutants were elucidated using nuclear magnetic resonance spectroscopy and mass spectrometry. The capped core of the wapR mutant was found to be truncated and lacked ␣-1,3-L-Rha. In contrast, uncapped core OS from the migA mutant lacked ␣-1,6-L-Rha. These results provide evidence that WapR is the ␣-1,3-rhamnosyltransferase, while MigA is the ␣-1,6-rhamnosyltransferase.Pseudomonas aeruginosa is an important gram-negative opportunistic pathogen capable of producing virulence determinants, including secreted toxins and enzymes, as well as cell surface structures, such as adhesins, flagella, and lipopolysaccharide (LPS) (18). LPS is a glycolipid composed of three distinct regions: lipid A, core oligosaccharide, and O antigen. P. aeruginosa produces two forms of O antigen, which are the homopolymeric A band and the heteropolymeric B band (15,21). The gene clusters associated with the biosynthesis of Aband and B-band O antigen in P. aeruginosa have been well characterized (for reviews, see references 18 and 22). In contrast, relatively little is known about the biosynthesis of the core oligosaccharide (OS).The P. aeruginosa core OS, like that of other gram-negative bacteria, can be conceptually subdivided into the inner core and the outer core (Fig. 1). The inner core is highly conserved and contains two L-glycero-D-manno-heptose (Hep) residues and two 3-deoxy-D-manno-octulosonic acid (Kdo) residues; the genetics and enzymology of this region are the best understood to date (14). The outer core is composed of four D-glucose (D-Glc) residues, one L-rhamnose (L-Rha) residue, and one N-(L-alanyl)-D-galactosamine residue, and its biosynthesis is not fully understood. The structures of the core OS of several different serotypes, clinical isolates, and rough mutants of P. a...

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Section: Discussionsupporting

confidence: 80%

mentioning

confidence: 67%

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Functional Characterization of MigA and WapR: Putative Rhamnosyltransferases Involved in Outer Core Oligosaccharide Biosynthesis ofPseudomonas aeruginosa

et al. 2008

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“…The S-type LPS is involved in nonmucoid and in early stage of P. aeruginosa infection in CF patients, whereas the R-type LPS is associated with mucoid and late stage of P. aeruginosa infection in CF patients [139]. The O-antigen is immunogenic in the host for the induction of protective antibodies, whereas lipid A is the core endotoxic component for induction of inlammatory responses [138]. More than 20 serotypes of O-antigens have been identiied [138].…”

Section: Antibody and Vaccine Development Against Lpsmentioning

confidence: 99%

“…LPS has two types, smooth or S-type and rough or R-type. S-type LPS consists of O-polysaccharide (O-antigen) repeats linked with a core-conserved oligosaccharide and a lipid A moiety, while R-type LPS lacks O-antigen and only contains the core oligosaccharide [138]. The S-type LPS is involved in nonmucoid and in early stage of P. aeruginosa infection in CF patients, whereas the R-type LPS is associated with mucoid and late stage of P. aeruginosa infection in CF patients [139].…”

Section: Antibody and Vaccine Development Against Lpsmentioning

confidence: 99%

Physiology and Pathology of Multidrug-Resistant Bacteria: Antibodies- and Vaccines-Based Pathogen-Specific Targeting

Zhang¹,

Su²,

Wu³

2017

Physiology and Pathology of Immunology

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Multidrug-resistant bacteria (MDR) are increasing rapidly and posing a global threat to mankind. Alternative strategies other than antibiotics have to be explored urgently. In this chapter, we review the current status of nonantibiotics strategies including antibodybased therapy and vaccine development for targeting Gram-positive strains (methicillinresistant Staphylococcus aureus and vancomycin-resistant Enterococcus faecium) and MDR Gram-negative strains (Acinetobacter baumannii and Pseudomonas aeruginosa). Biologicsbased clinical progress against these bacterial infections is updated.

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Prevotella denticola Lipopolysaccharide from a Cystic Fibrosis Isolate Possesses a Unique Chemical Structure

et al. 2016

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We report the first complete structural characterization of the lipopolysaccharide (LPS) from a cystic fibrosis (CF) clinical isolate of Prevotella denticola (B003V1S1X). Chemical, spectroscopic, and spectrometric analyses revealed a unique rough‐type LPS (LOS) structure. The structure has a highly negatively charged heptasaccharide core region containing hexoses, with the first two sugars, 3‐deoxy‐D‐manno‐oct‐2‐ulosonic acid (Kdo) and mannose, highly phosphorylated. Furthermore, the lipid A moiety has the typical structure for the genus Prevotella, and was also highly phosphorylated.

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Structures of the core oligosaccharide and O-units in the R- and SR-type lipopolysaccharides of reference strains ofPseudomonas aeruginosaO-serogroups

Cited by 60 publications

References 33 publications

Functional Characterization of MigA and WapR: Putative Rhamnosyltransferases Involved in Outer Core Oligosaccharide Biosynthesis ofPseudomonas aeruginosa

Functional Characterization of MigA and WapR: Putative Rhamnosyltransferases Involved in Outer Core Oligosaccharide Biosynthesis ofPseudomonas aeruginosa

Physiology and Pathology of Multidrug-Resistant Bacteria: Antibodies- and Vaccines-Based Pathogen-Specific Targeting

Prevotella denticola Lipopolysaccharide from a Cystic Fibrosis Isolate Possesses a Unique Chemical Structure

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