Bacterial Lipopolysaccharides: Relationship of Structure and Conformation to Endotoxic Activity, Serological Specificity and Biological Function

Rietschel, Ernst Theodor; Brade, Lore; Schade, Ulrich; Seydel, Ulrich; Zähringer, Ulrich; Brandenburg, Klaus; Helander, Ilkka M.; Holst, Otto; Kondo, Seiichi; Kuhn, H M

doi:10.1007/978-1-4757-5140-6_5

Cited by 54 publications

(40 citation statements)

References 8 publications

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“…The average length of the O chain in HF-LPS was less than one repeat. The revelation of almost one mol of direct Morgan-Elson reactivity in parallel with the loss of phosphate is as expected since the position C-1 of the reducing GIcN in S. typhimurium lipid A is known to carry a phosphate substituent [3].…”

Section: Resultssupporting

confidence: 71%

“…Different bacterial genera and species elaborate lipopolysaccharides which are compositionally different. A characteristic feature of LPS is that they always contain phosphorus; this is found linked to both lipid A and to the core oligosaccharide region [1][2][3]. The detection and quantitation of 3-deoxy-D-manno-octulosonic acid (dOclA), which links the core oligosaccharide to lipid A, is hampered by phosphate substituents [4].…”

Section: Introductionmentioning

confidence: 99%

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Cleavage of the O antigen 4,5,12 of Salmonella typhimurium by hydrofluoric acid

Helander

Kitunen

1989

FEBS Letters

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The effect of hydrofluoric acid (aqueous 48% HF) upon different lipopolysaccharides (LPS) was studied, employing conditions (48 h at + 4°C) that are commonly used to dephosphorylate LPS. From the LPS of Salmonella typhimurium having the O antigen 4,5,12 almost all of the O-antigenic sugars (Abe, Gal, GIc, Man, Rha) were liberated in dialysable form, whereas the saccharide chains of Salmonella LPS with O antigen 6,7 (Man, Glc, GIcNAc) were resistant to HF. The lability towards HF was shown to be due to the presence of the deoxysugar L-rhamnose in the saceharide backbone of the O antigen 4,5,12, since only Rha was found as the terminal sugar in the corresponding dialysable material. Hydrofluoric acid can thus be used to specifically cleave Rha-containing polysaceharides.

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Section: Resultssupporting

confidence: 71%

Section: Introductionmentioning

confidence: 99%

Cleavage of the O antigen 4,5,12 of Salmonella typhimurium by hydrofluoric acid

Helander

Kitunen

1989

FEBS Letters

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“…Earlier data show that the physicochemical state of lipopolysaccharides or lipid A greatly influences their biological [20] and serological [23] properties. Our present measurements reveal an inverse relationship between the chainmelting temperatue, t,, and the potency of inducing LTC, release (Fig.…”

Section: Discussionmentioning

confidence: 99%

Structural and physicochemical requirements of endotoxins for the activation of arachidonic acid metabolism in mouse peritoneal macrophages in vitro

Lüderitz¹,

Brandenburg²,

Seydel³

et al. 1989

European Journal of Biochemistry

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Lipopolysaccharides of different wild-type and mutant gram-negative bacteria, as well as synthetic and bacterial free lipid A, were studied for their ability to activate arachidonic acid metabolism in mouse peritoneal macrophages in vitro. It was found that lipopolysaccharides of deep-rough mutants of Saln~onellu minnesotu and Escherirhia coli (Re to Rc chemotypes) stimulated macrophages to release significant amounts of leukotriene C4 (LTC,) and prostaglandin E2 (PGE,). Lipopolysaccharides of wild-type strains (S. abortus equi, S.friedenau) only induced PGE2 and not LTC, formation. Unexpectedly, free bacterial and synthetic E. coli lipid A were only weak inducers of LTC, and PGE2 production. Deacylated Re-mutant lipopolysaccharide preparations were inactive. However, co-incubation of macrophages with both deacylated lipopolysaccharide and lipid A lead to the release of significant amounts of LTC, and PGE2, similar to those obtained with Re-mutant lipopolysaccharide. The significance of the lipid A portion of lipopolysaccharide for the induction of LTC4 was indicated by demonstrating that peritoneal macrophages of endotoxin-low-responder mice or of mice rendered tolerant to endotoxin did not respond with the release of arachidonic acid metabolites on stimulation with Re-mutant lipopolysaccharide and that polymyxin B prevented the Re-lipopolysaccharide-induced LTC, and PGE2 release. Physical measurements showed that the phase-transition temperatures of both free lipid A and S-form lipopolysaccharide were above 37 "C while those of R-mutant lipopolysaccharides were significantly lower (30 -35 "C). Thus, with the materials investigated, an inverse relationship between the phase-transition temperature and the capacity to elicit LTC4 production was revealed.In a previous paper we demonstrated that, under specified conditions, lipopolysaccharides of a Salmonella minnesota Re mutant (strain R595) are capable of inducing, in cultured mouse peritoneal macrophages, leukotriene C4 (LTC,) formation and release in a dose-and time-dependent manner [l]. Chemically, this lipopolysaccharide consists of lipid A carrying an a(2,4)-linked 3-deoxy-~-manno-octu~osonic acid (dOclA) disaccharide [2-41. It has recently been proven that the lipid A component represents the endotoxic principle of lipopolysaccharides and that it elicits many of the biological effects seen with endotoxins [5]. On the other hand, a number of studies indicate that also the polysaccharide portion possesses intrinsic biological activity or that it may modulate lipid A activity [6, 71. I t was, therefore, of interest to investigate the significance of the lipid A and the polysaccharide region in the lipopolysaccharide-induced formation of LTC4 in phagocytes.The present paper describes experiments involving lipopolysaccharides from wild-type bacteria (S-form lipopolysaccharides), rough mutants (R-form lipopolysaccharides) and free bacterial, as well as synthetic lipid A (preparation 506). Our results suggest that the physicochemical state of Abbreviations. P,/NaC...

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“…2 indicates, N. meningitidis lipid A belongs to the group of lipid A's possessing a symmetrical acylation pattern. This type of structure was first encountered in Chromobacterium violaceum (17,33,34) and, therefore, is referred to as a C. violaceum-type lipid A. Other members of this group are N. gonorrhoeae and Pseudomonas aeruginosa hexaacyl lipid A's (23,41).…”

Section: Methodsmentioning

confidence: 99%

Structural characterization of the lipid A component of pathogenic Neisseria meningitidis

et al. 1992

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The lipid A component of meningococcal lipopolysaccharide was structurally characterized by using chemical modification methods, methylation analysis, 31P nuclear magnetic resonance, and laser desorption mass spectroscopy. It was shown that Neisseria meningitidis lipid A consists of a 1,4'-bisphosphorylated 1(1'-+6)-linked D-glucosamine disaccharide (lipid A backbone), both phosphate groups being largely replaced by O-phosphorylethanolamine. This disaccharide harbors two nonsubstituted hydroxyl groups at positions 4 and 6', the latter representing the attachment site of the oligosaccharide portion in lipopolysaccharide. In addition, it is substituted by up to six fatty acid residues. In the major lipid A component, representing a hexaacyl species, the hydroxyl groups at positions 3 and 3' carry (R)-3-hydroxydodecanoic acid [12:0(3-OH)], whereas the amino groups at positions 2 and 2' are substituted by (R)-3-(dodecanoyloxy)tetradecanoic acid Meningococcal infections represent a serious problem for small children and young adults in many parts of the world, the lethality being high even in developed countries (1, 5, 6). The causative agent, Neisseria meningitidis, like other gramnegative bacteria, possesses an endotoxin complex located in the bacterial outer cell membrane. Endotoxin (lipopolysaccharide [LPS]) is known to cause or to contribute to the severe symptoms of meningitis, culminating in irreversible shock (37). Meningococci vary in the ability to release endotoxin-containing outer membrane vesicles into the surroundings, both in vitro and in vivo (7). Recent studies performed with N. meningitidis isolates from the nasopharynx of carriers and from cerebrospinal fluid and blood samples from patients with meningococcal infection demonstrated that overproduction and liberation of endotoxin are strongly associated with pathogenicity, regardless of the serological group or serotype of the strain (8, 9).Lipid A represents the principal structural component responsible for the harmful biological activities of LPS (32). To understand the structural parameters required for the expression of endotoxic activity, it appears important to know the precise chemical structure of the lipid A component of pathogenic gram-negative bacteria such as N. meningitidis. The present paper elucidates the primary structure of the lipid A component of N. meningitidis isolated from a strain (M986-NCV1) expressing the serogroup B LPS. It will be shown that this lipid A shares certain structural features with the well-studied enterobacterial lipid A but that it differs significantly in the substitution of the hydrophobic backbone and in the nature and locations of acyl chains. * Corresponding author. MATERIALS AND METHODSMicroorganism and LPS. N. meningitidis M986-NCV1, a nonencapsulated variant of the group B strain M986, was obtained from the culture collection of the Center for Biologics Evaluation and Research. This strain was grown overnight in tryptic soy broth (Difco Laboratories, Detroit, Mich.) in 2.8 liters of baffled Fernbach ...

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Bacterial Lipopolysaccharides: Relationship of Structure and Conformation to Endotoxic Activity, Serological Specificity and Biological Function

Cited by 54 publications

References 8 publications

Cleavage of the O antigen 4,5,12 of Salmonella typhimurium by hydrofluoric acid

Cleavage of the O antigen 4,5,12 of Salmonella typhimurium by hydrofluoric acid

Structural and physicochemical requirements of endotoxins for the activation of arachidonic acid metabolism in mouse peritoneal macrophages in vitro

Structural characterization of the lipid A component of pathogenic Neisseria meningitidis

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