RELATION OF STRUCTURE TO FUNCTION IN BACTERIAL O‐ANTIGENS. V. NATURE OF ACTIVE SITES IN ENDOTOXIC LIPOPOLYSACCHARIDES OF <i>SERRATIA MARCESCENS</i>*

Tripodi, D; Nowotny, Alois

doi:10.1111/j.1749-6632.1966.tb52392.x

Cited by 72 publications

(45 citation statements)

References 25 publications

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“…Changes in mouse lethality associated with alkaline hydrolysis correlates with changes in molecular symmetry. Molecular dyssemmetry, postulated to include unfolding and swelling of the molecule, increases progressively between the 3rd and the 8th hr (25). Changes associated with alkaline hydrolysis which have been shown to occur in other assay systems are in the same direction as the time-dependent changes demonstrated here in a cell culture system.…”

Section: Resultssupporting

confidence: 58%

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The effect of bacterial products on synovial fibroblast function: hypermetabolic changes induced by endotoxin

Buckingham¹,

Castor²

1972

J. Clin. Invest.

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Section: Resultssupporting

confidence: 58%

“…Pyrogenicity is not completely eliminated by such treatment. Lethality for mice, although not decreased in the first 3 hr, is virtually eliminated at 6-8 hr (25). Changes in mouse lethality associated with alkaline hydrolysis correlates with changes in molecular symmetry.…”

Section: Resultsmentioning

confidence: 92%

The effect of bacterial products on synovial fibroblast function: hypermetabolic changes induced by endotoxin

Buckingham¹,

Castor²

1972

J. Clin. Invest.

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“…The results also show that mere solubilization of lipid A by the aid of triethylamine [20] or pyridine [19] does not render lipid A anticomplementary. The results suggest that functional groups in lipid A must be exposed, which are blocked in aggregates of lipid A and which become available for interaction with complement in complexes with suitable carriers, where lipid A is present in a favourable conformation (see also [22,23]). A following paper will deal in more detail with the toxicity of lipid A carrier complexes.…”

Section: Mouse Lethality Testsmentioning

confidence: 99%

Interaction of Lipopolysaccharides and Lipid A with Complement

Galanos¹,

Rietschel²,

Lüderitz³

et al. 1971

European Journal of Biochemistry

262

156

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A number of lipopolysaccharides derived from Salmonella and Escherichia coli S and R mutant strains were tested for toxicity and anticomplementary activity in the absence of added antiserum. Although all preparations were toxic, only a few exhibited high anticomplementary activity, while others proved to be of low or negligible activity. It was found that isolated lipid A from both active and inactive lipopolysaccharides was strongly anticomplementary as well as toxic, when made water-soluble with the aid of suitable carriers. Treatment of R form lipopolysaccharide with Mg2+ or Ca2+ led to complete precipitation of the lipopolysaccharide with consequent loss of toxicity and anticomplementary activity. This treatment had practically no effect on the anticomplementary activity and toxicity of S form lipopolysaccharides. When lipopolysaccharide, after reaction with complement, was reisolated and pursed, the resulting preparation was found to be non-toxic and of negligible anticomplementary activity. No detectable alterations in either the sugar or the fatty acid composition could be detected. The only significant change was the loss of solubility in water. Treatment of the reisolated lipopolysaccharide with EDTA completely restored solubility in water, toxicity, and anti-complementary activity.It has been known for a long time [l] that the cell wall lipopolysaccharides of gram-negative bacteria exhibit anticomplementary activity. Recently, Mergenhagen et al. [2,3] have shown that complement-endotoxin interaction leads to loss of terminal complement activity with simultaneous appearance of lesions on the lipopolysaccharide molecule as judged by electron microscopy. I n experiments in vitro, it could also be demonstrated that complement-endotoxin interaction leads to certain biological activities such as the generation of anaphylatoxin, chemotactic factors and the activation of the clotting system [3,4]. Since these phenomena are also observed after injection of endotoxin in animals, it has been suggested [3] that some of the endotoxic activities of lipopolysaccharides are mediated through the activation of the complement system. The above studies were primarily concerned with the role of complement in complement-endotoxin interactions and the biological significance of this phenomenon.In this paper the emphasis is directed mainly towards the role and fate of the lipopolysaccharide in reactions involving complement, and in particular to the region of the molecule responsible for anticomplementary activity. Previous studiea have shown that for anticomplementary activity [2] and for toxicity the 0-spec& chains of lipopolysaccharides as well as part of their core polysaccharide are not necessary; i. e. R form lipopolysaccharides conUnwncal Abbreviation. KDO, 2-keto-3-deoxyoctonate.taining lipid A and 2-keto-3-deoxyoctonate (KDO) as the main constituents can be as active as the parent S form lipopolysaccharides. The role of lipid A in these reactions became thus more apparent.In the present study further evidence will b...

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“…Moreover, Niwa et al (14) and Tripodi et al (24) reported that the molecular weight of endotoxin was reduced when treated with alkali. These results suggest changes in aggregating form of the endotoxin.…”

Section: Discussionmentioning

confidence: 99%

“…One of the first modifications used was alkaline hydrolysis (20), which greatly reduced in vivo lethality (4, 5, 13, 14, 24), pyrogenicity (14, 23), and antitumor activity (4, 5, 23). Alkaline hydrolysis also cleaves the O-ester-linked fatty acids from the lipid A moiety (16), and reduces its particle size as determined by molecular weight and sedimentation velocity (12,14,24).…”

mentioning

confidence: 99%

Electron Microscopic Studies of Endotoxins Treated with Alkaline and Acid Reagents

Amano

Fukushi

1984

Microbiology and Immunology

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Endotoxins extracted from Salmonella minnesota wild strain and R mutants (Ra to Re) were treated with two different alkaline reagents. Treatment with diluted sodium hydroxide, which caused partial removal of O‐ester linked fatty acids, changed the ultrastructures of endotoxins from an onion‐like structure to monolayer particles (approximately 100 Å in diameter) except for endotoxic glycolipids from Rd2 and Re mutants which showed mixed ultrastructures of untreated and treated endotoxins. Treatment with alkaline hydroxylamine, which caused liberation of all O‐ester linked fatty acids, changed the ultrastructures of all endotoxins to monolayer particles. The results suggested that the ultrastructures of alkaline‐treated endotoxins were dependent on the degree of their hydrophobicity. On the other hand, the micrographs of acid treated endotoxins did not show a constant structure because of the high hydrophobicity.

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RELATION OF STRUCTURE TO FUNCTION IN BACTERIAL O‐ANTIGENS. V. NATURE OF ACTIVE SITES IN ENDOTOXIC LIPOPOLYSACCHARIDES OF *SERRATIA MARCESCENS**

Cited by 72 publications

References 25 publications

The effect of bacterial products on synovial fibroblast function: hypermetabolic changes induced by endotoxin

The effect of bacterial products on synovial fibroblast function: hypermetabolic changes induced by endotoxin

Interaction of Lipopolysaccharides and Lipid A with Complement

Electron Microscopic Studies of Endotoxins Treated with Alkaline and Acid Reagents

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RELATION OF STRUCTURE TO FUNCTION IN BACTERIAL O‐ANTIGENS. V. NATURE OF ACTIVE SITES IN ENDOTOXIC LIPOPOLYSACCHARIDES OF SERRATIA MARCESCENS*

Cited by 72 publications

References 25 publications

The effect of bacterial products on synovial fibroblast function: hypermetabolic changes induced by endotoxin

The effect of bacterial products on synovial fibroblast function: hypermetabolic changes induced by endotoxin

Interaction of Lipopolysaccharides and Lipid A with Complement

Electron Microscopic Studies of Endotoxins Treated with Alkaline and Acid Reagents

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Product

Resources

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RELATION OF STRUCTURE TO FUNCTION IN BACTERIAL O‐ANTIGENS. V. NATURE OF ACTIVE SITES IN ENDOTOXIC LIPOPOLYSACCHARIDES OF *SERRATIA MARCESCENS**