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

Boll, Matthias; Radziejewska‐Lebrecht, Joanna; Warth, Christoph; Krajewska-Pietrasik, D; Mayer, Hubert

doi:10.1111/j.1574-695x.1994.tb00460.x

Cited by 41 publications

(37 citation statements)

References 29 publications

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“…The possibility of generating pure 13 C-labeled Kdo 2 -Lipid A, as described above, should greatly facilitate NMR solution studies of complexes with MD2, or CD14. Heptose-deficient "Re" LPS from Salmonella minnesota, which is often used to stimulate immune cells, would not be suitable for such biophysical studies, because of its considerable micro-heterogeneity with regard to the number of acyl chains and polar substituents (48,61,62).…”

Section: Discussionmentioning

confidence: 99%

Kdo2-Lipid A of Escherichia coli, a defined endotoxin that activates macrophages via TLR-4

Raetz

Garrett

Reynolds

et al. 2006

Journal of Lipid Research

209

248

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The LIPID MAPS Consortium (www.lipidmaps. org) is developing comprehensive procedures for identifying all lipids of the macrophage, following activation by endotoxin. The goal is to quantify temporal and spatial changes in lipids that occur with cellular metabolism and to develop bioinformatic approaches that establish dynamic lipid networks. To achieve these aims, an endotoxin of the highest possible analytical specification is crucial. We now report a large-scale preparation of 3-deoxy-D-manno-octulosonic acid (Kdo) 2 -Lipid A, a nearly homogeneous Re lipopolysaccharide (LPS) sub-structure with endotoxin activity equal to LPS. Kdo 2 -Lipid A was extracted from 2 kg cell paste of a heptose-deficient Escherichia coli mutant. It was purified by chromatography on silica, DEAE-cellulose, and C18 reverse-phase resin. Structure and purity were evaluated by electrospray ionization/mass spectrometry, liquid chromatography/mass spectrometry and 1 H-NMR. Its bioactivity was compared with LPS in RAW 264.7 cells and bone marrow macrophages from wild-type and toll-like receptor 4 (TLR-4)-deficient mice. Cytokine and eicosanoid production, in conjunction with gene expression profiling, were employed as readouts. Kdo 2 -Lipid A is comparable to LPS by these criteria. Its activity is reduced by . The LIPID MAPS consortium is developing quantitative methods for evaluating the composition, biosynthesis, and function of all macrophage lipids (1). These amphipathic substances not only are structural components of biological membranes but also play important roles in the pathophysiology of inflammation, atherosclerosis, and growth control. Additional lipid functions should emerge from the comprehensive analysis of macrophage lipids. Electrospray ionization/mass spectrometry (ESI/MS) (2, 3), coupled with prefractionation methods like reversephase liquid chromatography (LC), is being applied systematically to set the stage for the seamless integration of lipid metabolism into the broader fields of genomics, proteomics, and systems biology. To facilitate this endeavor, LIPID MAPS has introduced a new comprehensive classification system for biological lipids, amenable to computer-based data processing and substructure comparison (4). The eight LIPID MAPS categories are 1) fatty acyls, 2) glycerolipids, 3) glycerophospholipids, 4) sphingolipids, 5) sterol lipids, 6) prenol lipids, 7) saccharolipids,

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Section: Discussionmentioning

confidence: 99%

Kdo2-Lipid A of Escherichia coli, a defined endotoxin that activates macrophages via TLR-4

Raetz

Garrett

Reynolds

et al. 2006

Journal of Lipid Research

209

248

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“…The lipid A moiety of lipopolysaccharide of certain Gramnegative bacteria can be modified with the positively charged sugar L-Ara4N (58,(62)(63)(64)(65)(66) in a regulated manner (26,28,55). By masking of the negative phosphate groups of lipid A, the L-Ara4N residue prevents binding of cationic antimicrobial peptides and polymyxin to the outer membrane, rendering the bacteria resistant to killing by these substances (30 -32, 37).…”

Section: Discussionmentioning

confidence: 99%

Accumulation of a Polyisoprene-linked Amino Sugar in Polymyxin-resistant Salmonella typhimurium andEscherichia coli

Trent¹,

Ribeiro²,

Doerrler³

et al. 2001

Journal of Biological Chemistry

135

208

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Polymyxin-resistant mutants of Escherichia coli andPolyisoprene-linked sugars function as donor substrates for many types of glycosyltransferases (1). In eubacteria, undecaprenyl moieties (1, 2) serve as lipid carriers for sugar residues that are transferred to acceptors located outside of the cytoplasm, where sugar nucleotides are not available. Undecaprenyl diphosphate-sugars (generally oligosaccharides) are precursors for polymerization of O-antigen (3-6), enterobacterial common antigen (7,8), and peptidoglycan (1, 9 -11). Undecaprenyl phosphate-sugars (typically monosaccharides) are thought to be donors for processes that include bacteriophagemediated O-antigen conversion (12, 13), glycosylation of teichoic acids (14 -17), and biosynthesis of mycobacterial lipoglycans (18 -21). In eucaryotic cells, the structurally related dolichyl phosphate-sugars and dolichyl diphosphate-sugars (1, 22-24) play important roles in various stages of protein glycosylation and in the assembly of phosphatidylinositol-linked glycans.As demonstrated in the preceding article (25), a membranebound donor, proposed to be undecaprenyl phosphate-␣-LAra4N 1 (Fig. 1) 2 based upon bioinformatic considerations (26, 27), is required for the modification of lipid A with 4-amino-4-deoxy-L-arabinose (L-Ara4N) units in polymyxin-resistant mutants of Escherichia coli and Salmonella typhimurium. A novel L-Ara4N transferase, encoded by arnT (previously designated orf5 or pmrK) (28, 29), catalyzes L-Ara4N transfer to lipid A-like molecules in vitro when membranes of polymyxin-resistant mutants are employed as the source of the L-Ara4N donor (25). The formation of L-Ara4N and its transfer to lipid A are induced by activation of the transcription factor PmrA, which may occur by mutation (30 -32), by activation of PhoP (33, 34), or by exposure of cells to mildly acidic pH, ferric ions, or metavanadate (26,35,36). Attachment of the positively charged L-Ara4N moiety to lipid A is critical for resistance to the antibiotic polymyxin and to certain cationic antimicrobial peptides present inside phagocytic cells (37,38).We now report the purification and structural characterization of a novel, minor lipid that accumulates in polymyxinresistant mutants of E. coli and S. typhimurium. The purified lipid functions as a donor of L-Ara4N residues in the ArnTcatalyzed modification of lipid A in vitro. MALDI/TOF mass spectrometry and high resolution NMR spectroscopy strongly * This work was supported in part by National Institutes of Health Grants GM-51310 (to C. R. H. R.) and GM54882 (to R. J. C). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

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“…Polymyxins are cationic and show high affinity for negatively charged surfaces such as the lipopolysaccharide (LPS) in the outer membrane, and not surprisingly, microorganisms that are resistant to polymyxin have a modified LPS that binds less polymyxin due to a lower overall negative charge (53,55). This is primarily due to neutralization of the acidic phosphate on lipid A (3,24,25,36,56). For example, polymyxin-resistant mutants of Salmonella typhimurium and Escherichia coli have a higher substitution of the ester-linked phosphate group in the lipid A portion of the LPS by 4-amino-4-deoxy-L-arabinose and show larger amounts of 2-aminoethanol esterifying phosphates in the core oligosaccharide (24,29,54).…”

mentioning

confidence: 99%

Regulation of polymyxin resistance and adaptation to low-Mg2+ environments

1997

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The PmrA-PmrB two-component system of Salmonella typhimurium controls resistance to the peptide antibiotic polymyxin B and to several antimicrobial proteins from human neutrophils. Amino acid substitutions in the regulatory protein PmrA conferring resistance to polymyxin lower the overall negative charge of the lipopolysaccharide (LPS), which results in decreased bacterial binding to cationic polypeptides and increased bacterial survival within human neutrophils. We have now identified three PmrA-activated loci that are required for polymyxin resistance. These loci were previously shown to be necessary for growth on low-Mg

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

Cited by 41 publications

References 29 publications

Kdo2-Lipid A of Escherichia coli, a defined endotoxin that activates macrophages via TLR-4

Kdo2-Lipid A of Escherichia coli, a defined endotoxin that activates macrophages via TLR-4

Accumulation of a Polyisoprene-linked Amino Sugar in Polymyxin-resistant Salmonella typhimurium andEscherichia coli

Regulation of polymyxin resistance and adaptation to low-Mg2+ environments

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