Temperature-dependent incorporation of 4-amino-l-arabinose in lipid A of distinct Gram-negative bacteria

Lakshmi, Soundarajan; Bhat, U. Ramadas; Wartenberg, Klaus; Schlecht, S; Mayer, Hubert

doi:10.1111/j.1574-6968.1989.tb03493.x

Cited by 9 publications

(6 citation statements)

References 21 publications

(10 reference statements)

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“…However, antibiotic sensitivity in vivo is a very complex process (4,5,9,27,48) and may not be a function only of the membrane permeation property. Besides, the importance of fatty acyl chains attached to the lipid A moiety of LPS in membrane barrier function cannot be ignored (1,28,39,40,41,49,52). The fatty acyl chains of lipid A in Salmonella, Proteus, and Chromobacterium spp.…”

Section: Discussionmentioning

confidence: 99%

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%

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

1994

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“…Lipid A was precipitated from isolated LPS by hydrolysis either with 0.1 M sodium acetate buffer (pH 4.5, 2.5 h, 100°C) [19], or with 0.02 M HCI (10 min, 100°C) [16]. After centrifugation (10000 x g, 30 min), the precipitates were washed twice with water (20°C and 40°C), then with ice-cold ethanol and finally with ice-cold acetone.…”

Section: Lipid a Preparationmentioning

confidence: 99%

“…Lipid A bound L-Arap4N was liberated from LPS (5 mg) by hydrolysis with 0.02 M HCI for 10 min at 100°C) [16]. After centrifugation (10 000 × g, 10 min) the supernatant was passed over a 331 filter (Millipore, 0.2 /zm) and lyophilized.…”

Section: Detection Of Lipid a Bound 4-amino-4-deoxy-larabinosementioning

confidence: 99%

“…In the present study the lipid A/Kdo regions of strains with very similar lipid A structures were investigated which, however, showed natural or temperature-dependent differences in their incorporation of L-Arap4N [16]. The influence of the amount of L-Arap4N on both, the minimal inhibitory concentration of polymyxin B on the mutant strains and the affinity of pmxB to isolated LPS were investigated.…”

Section: Introductionmentioning

confidence: 98%

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4-Amino-4-deoxy-l-arabinose in LPS of enterobacterial R-mutants and its possible role for their polymyxin reactivity

Boll

Radziejewska‐Lebrecht²,

Warth

et al. 1994

FEMS Immunology &Amp; Medical Microbiology

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The content of 4-amino-4-deoxy-L-arabinopyranose (L-Arap4N) and the phosphate substitution pattern of the LPS of various strains from Salmonella minnesota, Yersinia enterocolitica and Proteus mirabilis was determined by GC/MS, HPLC and 31P-NMR. These data allowed us to examine the possible role of these components for the polymyxin B-binding capacity of LPS and for the minimal inhibiting concentration (MIC) and the minimal bactericidal concentration (MBC) of polymyxins B and E towards the respective R-mutants. Contrary to other investigated Re-, Rd- and Rc-mutants of S. minnesota, strain R595 (Re-mutant) showed about a 90% substitution of the ester-linked phosphate-group with L-Arap4N, whereas the L-Arap4N content of the other S. minnesota strains amounted to 17-25%. Neither the binding capacity of LPS to polymyxin B, determined by a bioassay, nor the MIC- and MBC-values of the R-mutants were significantly affected by this alteration. Similar results were obtained after using the temperature-dependent changes in the L-Arap4N-content and phosphate substitution pattern of Y. enterocolitica 75R. In order to explore the relevant polymyxin B binding site, lipid A samples with or without substitution of their ester-linked phosphate group were prepared and subjected to the polymyxin-binding assay. The results obtained so far indicated that the inner core bound L-Arap4N, detected in all resistant strains investigated, may play a decisive role in the decreased binding of polymyxin B, responsible for the bacterial resistance towards polymyxin(s).

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“…Most striking is the almost complete downregulation of Oantigen expression in bacteria grown at 37°C; another is the modification of the lipid A structure with respect to its substitutions such as phosphate, aminoarabinose and acyl chains (Lakshmi et al, 1989;Aussel et al, 2000). Details of the regulatory mechanisms are not known.…”

Section: Temperature and Other Environmental Stimulimentioning

confidence: 94%

Y. enterocolitica and Y. pseudotuberculosis

Carniel¹,

Autenrieth²,

Cornelis³

et al. 2006

The Prokaryotes

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Temperature-dependent incorporation of 4-amino-l-arabinose in lipid A of distinct Gram-negative bacteria

Cited by 9 publications

References 21 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

4-Amino-4-deoxy-l-arabinose in LPS of enterobacterial R-mutants and its possible role for their polymyxin reactivity

Y. enterocolitica and Y. pseudotuberculosis

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