31P nuclear magnetic resonance spectroscopy of lipopolysaccharides from pseudomonas aeruginosa

Horton, Derek; Riley, David

doi:10.1016/0005-2736(81)90103-6

Cited by 15 publications

(15 citation statements)

References 17 publications

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“…Phosphorylation of LPS is not a unique feature of P. syringae since a mature LPS is known to contain 2 to 6% phosphate and this figure is generally on the higher side in Pseudomonas species (13,51). Therefore, radiolabelling of LPS with 32p; in vivo or with [y-32P]ATP or -GTP in vitro is possible during its biosynthesis.…”

Section: Discussionmentioning

confidence: 99%

“…The structure of the 0 polysaccharide is most variable, and the 0 polysaccharide may contain various unusual sugars (32,38). The other striking feature of LPS is the presence of a high level of phosphorus (2 to 5.6%), which is attached either to the core sugars (such as 2-keto-3-deoxyoctonic acid and heptose) or to GlcNAc of lipid A as ethanolamine phosphate, ethanolamine PPi, PP1, or simply phosphate (13,25,38,51).Although much information about LPS composition and structure has accumulated during the last 25 years, very little is known about the physiological function of LPS. LPS have been studied as toxins and virulence factors of gram-negative bacteria that cause pathogenesis in animals and plants (19,32).…”

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Phosphorylation of lipopolysaccharides in the Antarctic psychrotroph Pseudomonas syringae: a possible role in temperature adaptation

1994

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Phosphorylation of lipopolysaccharide (LPS) from a psychrotrophic bacterium, Pseudomonas syringae, from Antarctica was studied by using sucrose gradient-separated membrane fractions. The bacterium was found to possess an LPS kinase which could phosphorylate more LPS postsynthetically at higher temperatures. The phosphorylation was low at a lower temperature and was also found to occur in vivo. After phosphorylation of LPS in vitro, it was found that the major part of the radioactivity (>85%) was associated with the core oligosaccharide region of the LPS. The phosphate groups of this region are probably involved in the binding of metal ions, which could be removed by EDTA. The cells grown at the lower temperature probably contained fewer divalent cations because of the smaller amount of phosphate and thereby were more sensitive to EDTA. The cells were also more sensitive to cationic antibiotics at the lower temperature. A possible role of this differential phosphorylation of LPS in modulating the function of the outer membrane as a permeability barrier in the psychrotroph is discussed.The outermost layer of the outer membrane bilayer of gram-negative bacteria is composed of lipopolysaccharides (LPS) which contain a hydrophobic moiety, lipid A, and a polysaccharide moiety consisting of the so-called core oligosaccharide and 0 polysaccharide. The chemical structures of lipid A from various bacteria such as Escherichia coli, Pseudomonas aeruginosa, Proteus mirabilis, Salmonella minnesota, Salmonella typhimurium, Shigella sonnei, Yersinia pestis, Er-winia carotovora, and Haemophilus ducreyi are remarkably identical in that they have a backbone of 3-1',6-glucosamine (GlcNAc) (19,32). Heterogeneity of lipid A arises because of variation in the fatty acid composition, which appears to vary depending on the bacterial strain and growth conditions (6,11,21,25,39,40,41,49,52). The composition of the core oligosaccharide is also variable; this oligosaccharide is generally composed of 2-keto-3-deoxyoctonic acid, heptose, glucose, and galactose. The structure of the 0 polysaccharide is most variable, and the 0 polysaccharide may contain various unusual sugars (32,38). The other striking feature of LPS is the presence of a high level of phosphorus (2 to 5.6%), which is attached either to the core sugars (such as 2-keto-3-deoxyoctonic acid and heptose) or to GlcNAc of lipid A as ethanolamine phosphate, ethanolamine PPi, PP1, or simply phosphate (13,25,38,51).Although much information about LPS composition and structure has accumulated during the last 25 years, very little is known about the physiological function of LPS. LPS have been studied as toxins and virulence factors of gram-negative bacteria that cause pathogenesis in animals and plants (19,32). However, the function of LPS with respect to the physiology of the bacteria is poorly understood. In bacteria, LPS has been implicated in providing a kind of permeation barrier for hydrophobic compounds to move across the membrane. The evidence for this has come from the study of...

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

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Phosphorylation of lipopolysaccharides in the Antarctic psychrotroph Pseudomonas syringae: a possible role in temperature adaptation

1994

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“…The spectrum is dominated by pyrophosphate signals at about -9.7 and -4.5 ppm, but also contains three signals of similar intensity at +3.76, +3.39, and +3.14 ppm (all probably attributable to phosphomonoester residues), and a minor signal at -0.34 ppm. The latter signal was absent from the corresponding spectra of lipopolysaccharides from P. aeruginosa NCTC 1999 and NCIB 8626 [30], but a signal in this region has been reported [35] for the lipopolysaccharides from the Fisher immunotype strains of P. aeruginosa. Only phosphomonoester signals were detected in the 31P NMR spectrum of the core oligosaccharide isolated after mild acid hydrolysis of the lipopolysaccharide from strain NCTC 8505, during the course of which much phosphorus is released in the form of Pi and ethanolamine phosphates [19].…”

Section: Phosphate Residues In the Lipopolysaccharidementioning

confidence: 99%

The Lipopolysaccharide from Pseudomonas aeruginosa NCTC 8505

Tahara¹,

Wilkinson²

1983

European Journal of Biochemistry

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Structural studies have been carried out on the 0-specific fraction from the lipopolysaccharide of Pseudomonas aeruginosa NCTC 8505, Habs serotype 03. The 0-specific polysaccharide has a tetrasaccharide repeating-unit containing residues of L-rhamnose (Rha), 2-acetamido-2-deoxy-~-glucose (GlcNAc), 2-acetamido-2-deoxy-~-galacturonic acid (GalNAcA), and 2,4-diacetamid0-2,4,6-trideoxy-i~-glucose (BacNAc,). The following structure has been assigned to the repeating-unit :The parent lipopolysaccharide is a mixture of S, R, and SR species, and its high phosphorus content is partly due to the presence of triphosphate residues, as found for other lipopolysaccharides from P. aeruginosa. In addition to phosphorus, heptose, a 3-deoxyoctulosonic acid, and amide-bound alanine, the core oligosaccharide contains glucose, rhamnose, and galactosamine (molar proportions 3 : 1 : 1). The rhamnose and part of the glucose are present as unsubstituted pyranoside residues: other glucose residues are 6-substituted. We have previously reported general analyses of the lipopolysaccharide from P. aeruginosa NCTC 8505 [19], belonging to Habs serogroup 03, and identified the amino sugars present in the 0-specific fraction [20,21]. Here we describe the results of further studies of these products, including evidence for the structure of the 0-specific polysaccharide. MATERIALS AND METHODS Isolation and Fractionation of LipopolysaccharideMethods used for the batch culture of Pseudomonas aeruginosa NCTC 8505, the preparation of cell walls, and the Abbreviations. BacNAc,, 2,4-diacetamido-2,4,6-trideoxygluccse (NN-diacetylbacillosamine); GalNAcA, 2-acetamido-2-deoxygakacturonic acid; GlcNAc, 2-acetamido-2-deoxyglucose; Rha, rhamnose.

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“…Because MALDI-TOF analysis is not quantitative, differences in polymyxin resistance observed among mutant strains may be due to differences in aminoarabinose content. Alternatively, differences may be due to changes in labile modifications of lipid A (e.g., phosphoethanolamine) lost during sample preparation (41) or to changes in nonlipid A surface structures, such as proximal LPS core sugar phosphates (21) or LPS-associated lipoproteins (20). Despite these possibilities, these results indicate that the P. aeruginosa PmrAB system mediates the addition of aminoarabinose to lipid A and provides additional support for the importance of this outer membrane modification in the polymyxin resistance of gram-negative bacteria.…”

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PmrAB, a Two-Component Regulatory System of Pseudomonas aeruginosa That Modulates Resistance to Cationic Antimicrobial Peptides and Addition of Aminoarabinose to Lipid A

Moskowitz¹,

Ernst²,

2004

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Spontaneous polymyxin-resistant mutants of Pseudomonas aeruginosa were isolated. The mutations responsible for this phenotype were mapped to a two-component signal transduction system similar to PmrAB of Salmonella enterica serovar Typhimurium. Lipid A of these mutants contained aminoarabinose, an inducible modification that is associated with polymyxin resistance. Thus, P. aeruginosa possesses a mechanism that induces resistance to cationic antimicrobial peptides in response to environmental conditions. Cationic antimicrobial peptides (CAPs) are a widely conserved host defense mechanism of plants and animals. Their antimicrobial effects can be attributed to their amphipathic, detergent-like nature, which enables individual CAP molecules to interact with both anionic and hydrophobic components of the bacterial envelope. CAPs bind to lipopolysaccharide (LPS), a major component of the gram-negative cell surface, through interactions with phosphates and fatty acids of LPS core and lipid A moieties (31). These molecules cross the outer membrane and periplasm, disrupt the membrane potential of the inner membrane, and thereby cause cell death (25). In vertebrates, the CAPs that pathogens encounter at epithelial surfaces are a major component of innate immunity, an ancient system of host defense that is stimulated via receptors that recognize pathogen-associated molecular patterns (29).Pseudomonas aeruginosa is an opportunistic pathogen of humans that causes infections in those with host defense defects such as epidermal injury, immunodeficiency, and impaired epithelial clearance mechanisms. In the human host, P. aeruginosa is exposed to endogenous CAPs such as ␤-defensins (37) and cathelicidins (5) at epithelial surfaces. It may also encounter exogenous CAPs in this setting, when agents such as the polymyxins, acylated cyclic CAPs synthesized by the grampositive soil bacterium Bacillus polymyxa, are used as antibiotics. Since the discovery and initial clinical use of the polymyxins more than 50 years ago, both clinical (12, 23, 26) and experimental (7, 16, 32) P. aeruginosa polymyxin resistance has been reported. P. aeruginosa possesses proteases that can degrade some CAPs (35); in addition, physiological (or "adaptive") polymyxin resistance may occur in response to membrane stresses such as divalent cation limitation (7,13,27,30) and polymyxin exposure (9, 16, 36), the latter being associated with the modulation of lipid A fatty acid composition (9). The P. aeruginosa PhoPQ two-component system contributes to the induction of these resistance phenotypes; however, its role appears to be complex (13,27), and the potential roles of other regulatory systems related to PmrAB, a response regulatorsensor kinase pair that regulates polymyxin resistance in Salmonella enterica serovar Typhimurium (18, 34), have not been defined.Isolation of polymyxin-resistant mutants of P. aeruginosa. Conditions that physiologically induce polymyxin resistance have not been fully defined for P. aeruginosa and could involve multiple regulatory...

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31P nuclear magnetic resonance spectroscopy of lipopolysaccharides from pseudomonas aeruginosa

Cited by 15 publications

References 17 publications

Phosphorylation of lipopolysaccharides in the Antarctic psychrotroph Pseudomonas syringae: a possible role in temperature adaptation

Phosphorylation of lipopolysaccharides in the Antarctic psychrotroph Pseudomonas syringae: a possible role in temperature adaptation

The Lipopolysaccharide from Pseudomonas aeruginosa NCTC 8505

PmrAB, a Two-Component Regulatory System of Pseudomonas aeruginosa That Modulates Resistance to Cationic Antimicrobial Peptides and Addition of Aminoarabinose to Lipid A

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