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Query: UNIPROT:P43026 (lipopolysaccharide)
62,215 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We extracted an R-form lipopolysaccharide (LPS) by the phenol-water method from Klebsiella sp. strain LEN-111 (O3-:KI-) and followed the changes in ultrastructure of the LPS during the extraction procedure. When the LPS was obtained from the water phase of an extract by addition of 2 volumes of 10 mM MgCI2-ethanol, it consisted of membrane pieces with a hexagonal lattice structure with a lattice constant of 14 to 15 nm. The lattice structure of the LPS was disrupted into short rods with sodium dodecyl sulfate, but the same hexagonal lattice structure was again formed by precipitation with 2 volumes of 10 mM MgCI2-ethanol. The LPS preparation after two cycles of treatment by the phenol-water method, which contained no detectable amounts of proteins, kept an unaltered ability to form the hexagonal lattice structure. Extensive treatment with pronase and extraction with chloroform did not impair the ability of the LPS preparation to form the lattice structure. When the other salts, NaCI, CaCI2 or Zn(CH3COO)2, were used for precipitation of the LPS with ethanol in place of MgCI2, the LPS did not form the hexagonal lattice structure. However, if the LPS precipitated with NaCI-ethanol was converted to the magnesium salt form after it was electrodialyzed, it formed the same hexagonal lattice structure as the LPS precipitated with MgCI2-ethanol. From these results, it was concluded that the R-form LPS has the ability of in vitro self-assembly into a hexagonal lattice structure in the presence of Mg2+ without the help of other components such as proteins and free lipids from outer membrane.
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PMID:Formation of a hexagonal lattice structure by an R-form lipopolysaccharide of Klebsiella sp. 399 76

A lipopolysaccharide (LPS) was isolated from the Lyme disease spirochete by a modification of the hot phenol-water method. The material was composed of 45% carbohydrate, 8% protein, 44% lipid A, and 1% 3-deoxy-D-mannooctulosonic acid and accounted for approximately 1.5% of the cellular dry weight. The isolated LPS possessed several biologic activities characteristic of endotoxins. The LPS was pyrogenic for rabbits, mitogenic for human mononuclear cells and murine splenocytes, capable of clotting limulus lysate, and cytotoxic for murine macrophages. LPS extracted from Borrelia burgdorferi by the petroleum-ether:chloroform:liquid-phenol procedure was also characterized. The results show that the Lyme disease spirochete contains a hitherto unknown LPS that is biologically active in vitro, and the expression of such activities in vivo may play an important role in the pathogenesis of Lyme disease. Some of the clinical manifestations of other spirochetal disease may be explained by similar endotoxins in those organisms. To our knowledge this is the first report of an LPS extracted from a spirochete that is known to be a human pathogen.
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PMID:Chemical and biologic characterization of a lipopolysaccharide extracted from the Lyme disease spirochete (Borrelia burgdorferi). 400 83

The structure of the polysaccharide part of the lipopolysaccharide from Bacteroides fragilis NCTC 9343 has been determined using sugar and methylation analysis as the principal tools. Phenol--water extraction followed by a phenol--chloroform--light petroleum extraction yielded a lipopolysaccharide suitable for structural analysis. Analysis of sugars using alditol acetates showed that the polysaccharide contained L-rhamnose, D-galactose and D-glucose in the approximate molar ratios of 1:5:1. After weak acid hydrolysis, two polysaccharide fractions were isolated by gel permeation chromatography: PSI and PSII with the sugar molar ratios 1:5:1 and 1:2:1 respectively. Chromium trioxide oxidation revealed that all galactosyl residues have the beta configuration, and that the rhamnosyl and glucosyl residues have the alpha configuration. From methylation analysis of lipopolysaccharide and the PS I and PS II fractions the following structures could be deduced.
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PMID:Structural studies of the polysaccharide part of the cell wall lipopolysaccharide from Bacteroides fragilis NCTC 9343. 402 52

A modified Limulus amebocyte lysate (LAL) test was developed which quantifies the inhibition associated with lipopolysaccharide (LPS) in serum and plasma. The assay utilized the LR50 value to determine relative inhibition. The LR50, measured in ng/ml, represented the concentration of endotoxin needed to elicit a turbidimetric response equal to 50% of the maximum above the control (no added endotoxin). The LR50 concept was used to compare plasma and serum from a normal donor population. Plasma LR50 values ranged from 115 to 5000 ng/ml, while serum samples ranged from 48 to 615 ng/ml. Endotoxin in saline measured in a similar manner yielded values averaging 0.07 ng/ml (LAL sensitivity). Various means to remove inhibition were also tested by this method. Heating, dilution, and chloroform extraction of serum were all found to be efficient in removing inhibition. Although no direct attempt was made to elucidate the biochemical nature of the inhibitor(s) present in serum and plasma, the results presented were consistent with other studies and indicate the involvement of a heat labile component(s). The LR50 may provide a tool for the elaboration of these components. It also was suggested that LR50 may be the best in vitro indicator of the overall ability of serum or plasma to neutralize endotoxin.
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PMID:Quantification of endotoxin inhibition in serum and plasma using a turbidimetric LAL assay. 404 3

An R-form lipopolysaccharide (LPS) extracted from Klebsiella strain LEN-111 (O3-:K1-) by the phenol-chloroform-petroleum ether method was compared with that extracted by the phenol-water method in the ability to form a hexagonal assembly. The LPS which was extracted by the phenol-water method and dialyzed against tap water to remove phenol showed ribbon-like structures, and it formed a hexagonal lattice structure with a lattice constant of 14.5 +/- 0.3 nm when it was precipitated by addition of two volumes of 10 mM MgCl2-ethanol. The LPS which was extracted by the phenol-chloroform-petroleum ether method and lyophilized consisted of ribbon-like structures and their fragments and it often formed small pieces of a hexagonal lattice, although the LPS before lyophilization did not form such a lattice. When the LPS extracted by the phenol-chloroform-petroleum ether method was precipitated by addition of two volumes of 10 mM MgCl2-ethanol, it formed essentially the same hexagonal lattice structure as that formed by the LPS extracted by the phenol-water method. From these results it is concluded that the ability of the LPS to form a hexagonal lattice structure does not depend upon the method of its extraction from bacterial cells.
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PMID:Formation of a hexagonal lattice structure by an R-form lipopolysaccharide of Klebsiella: comparative study of preparations extracted by the phenol-water and the phenol-chloroform-petroleum ether methods. 409 70

Of the numerous members of the family Enterobacteriaceae only a few strains, notably Escherichia coli O14 and R mutants of the E. coli R1-core type, engender antibodies against the common enterobacterial antigen (CA) following immunization of rabbits with heated suspensions or culture supernatants; other members produce nonimmunogenic CA of identical serological specificity. The biochemical basis of the immunogenic properties of CA of the former strains was investigated by determining the relationship between the CA determinant and the lipopolysaccharide molecule. Lipopolysaccharides extracted from R mutants of the E. coli R1-core type or of E. coli O14 by the phenol-chloroform-petrol ether method contain the CA determinant, in contrast to extracts of other CA-producing R mutants. This is evident from the observation that only the former absorb CA antibodies, utilizing erythrocytes coated with alkali-treated lipopolysaccharide preparations. Based on the findings that CA of R mutants of E. coli R1-core type follows lipopolysaccharide during all purification steps and that alkali treatment increases its affinity for erythrocytes parallel to that of the lipopolysaccharide, it is concluded that the CA determinant either is part of the lipopolysaccharide molecule or is strongly complexed with it. It is suggested that this association between CA and the lipopolysaccharide of E. coli R1-core type and E. coli O14 accounts for the heat stability of the immunogenicity of CA of these unusual strains.
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PMID:Biochemical basis of the immunogenicity of the common enterobacterial antigen. 411 98

The endotoxin of a heptoseless mutant of Salmonella minnesota R595 was extracted with phenol-water. Most of this material was found distributed in the insoluble fraction of the extract. The results showed that the R595 endotoxin behaved as a lipid rather than as a lipopolysaccharide (LPS). The preparation, although it does not contain O-specific polysaccharides, does contain 2-keto-3-deoxyoctonic acid (KDO), hexosamine, and several other unidentified compounds. Therefore, the term "glycolipid" is used in this paper instead of lipopolysaccharide. The crude glycolipid fraction, which was soluble in a mixture of chloroform-methanol (8:2), was purified by a procedure including fractionation with organic solvents and by different-column chromatographic methods. Although a chromatographic fraction of the glycolipid showed homogeneity in most systems investigated, the presence of contaminants could not be excluded. Chemical analysis of the glycolipids showed the absence of hexoses and heptoses. Constituents which were found were hexosamine, KDO, fatty acids, and phosphorus, which showed a relatively simple chemical composition. Partial acidic hydrolysis of the glycolipid yielded hexosamine-phosphates, as described in "Lipid A" fractions of smooth LPS preparations. Thin-layer chromatography of the partially hydrolyzed glycolipid showed a pattern similar to "Lipid A" fractions of other strains. The biological activity of the glycolipid was at the same level as that of other gram-negative endotoxins. Pyrogenicity, Shwartzman reactivity, and chick embryo ld(50) values were as high or higher than those of purified Serratia marcescens endotoxin preparations, but mouse ld(50) measurements gave significantly lower results.
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PMID:Endotoxic glycolipid from a heptoseless mutant of Salmonella minnesota. 496 63

The identity of a heteropolysaccharide from cell walls of Mycobacterium tuberculosis H37Ra with Seibert's tuberculopolysaccharide I was demonstrated by thin-layer chromatography, chemical analysis, and antigenic tests. The polysaccharide of M. kansasii was shown to be identical with that of M. tuberculosis. Defatted cells were disintegrated by ultrasonic treatment in the presence of glass beads; cell walls were obtained by differential ultracentrifugation. Ethyl alcohol-precipitated carbohydrate extracts were analyzed for protein and nucleic acid; these impurities were removed. Tuberculopolysaccharide I from the mycobacterial culture filtrate is probably derived from a lipopolysaccharide of the cell wall, which is partially removed by chloroform in the intact state. Alkaline extraction releases additional polysaccharide, in varying degrees of association with cell wall murein.
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PMID:Chemical and serological relationships between the heteropolysaccharides of Mycobacterium tuberculosis and Mycobacterium kansasii. 496 49

Lysine limitation during growth of the lysine-requiring mutant of Escherichia coli 12408 resulted in the excretion of a complex containing 60% of lipopolysaccharide, 26% of extractable phospholipid and 11% of protein. The complex was obtained from the culture filtrate in yields of about 0.5g./l. by precipitation with chloroform or gel filtration; some purification steps are described. The greater part of the phospholipid consisted of phosphatidylethanolamine, which contained four main fatty acids; two monoenoic acids and a cyclopropane acid were esterified mainly in the beta-position, and a saturated acid was located mainly in the gamma-position. The protein component was relatively insoluble and contained an excess of acidic over basic amino acids and little cystine. The lipopolysaccharide resembled in composition the intracellular lipopolysaccharides from rough strains of E. coli. Both protein and lipopolysaccharide constituents of the complex were serologically active; the complex was less toxic than the purified lipopolysaccharide. In the electron microscope the complex showed a mixture of particles of various sizes and shapes. Rods and hollow spheroids (diameter 12-200mmu) were most common and resembled the particles previously found surrounding cells actively excreting the complex. The chloroform-precipitated material showed a tubular lamellar structure. Soluble lipopolysaccharide prepared from the complex also consisted of hollow spheres and rods.
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PMID:An extracellular lipopolysaccharide-phospholipid-protein complex produced by Escherichia coli grown under lysine-limiting conditions. 534 May 6

Phenol-soluble lipopolysaccharides were obtained from the interphase and phenol phase fractions of 44% aqueous phenol-extracted Citrobacter species. Upon detailed investigation of C. freundii 8090, the two lipopolysaccharide fractions were found to contain different amounts of lipid A, although qualitative composition was similar. Both contained lipid A, 2-keto-deoxyoctonic acid, heptose, phosphate, d-glucose, galactose, rhamnose, 2-acetamido-2 deoxy-d-glucose, 3-acetamido-3,6-dideoxy-d-glucose, O-acetyl, and trace amino acids. Partially purified phenol-phase lipopolysaccharide partitioned into the phenol-soluble phase when refractionated with 44% aqueous phenol, and was further found to be soluble in 88% phenol, 95% ethyl alcohol, and chloroform-methanol (2:1).
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PMID:Carbohydrate composition of the phenol-soluble lipopolysaccharides of Citrobacter freundii. 566 88


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