Structural studies on the O-specific side-chains of the cell-wall lipopolysaccharide from Salmonella typhimurium LT2

Hellerqvist, Carl G.; Lindberg, Bengt; Svensson, Sigfrid; Holme, Tord; Lindberg, Alf A.

doi:10.1016/s0008-6215(00)82139-4

Cited by 87 publications

(56 citation statements)

References 5 publications

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“…The sensitivity of the instrument could detect as a signif~cant peak 100 units of heptose or 2-acetamid0-2-deoxy-~-gluwse, both common to endotoxin core structure. A 1% endotoxin wntamination by a smooth LPS would contain at least 2% heptose and somewhat more amino sugar (7,16,17), or 783 area units. Because none was detected, then, based on the sensitivity of the instrument, there must be less than 100 ng endotoxin per 1000 pg of toxin if the former was present.…”

Section: Control For Gram-negative Contaminationmentioning

confidence: 99%

Studies on Group B β-Hemolytic Streptococcus. I. Isolation and Partial Characterization of an Extracellular Toxin

et al. 1981

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SummaryTo initiate an investigation into the biochemistry and mechanism of group B b-hemolytic Streptococcus virulence, bacterial cultures grown in suspension were centrifuged, and the bacteria and media were subjected to extensive fractionation. Each fraction was assayed for physiologic activity by repeated intravenous infusion into adult unanesthetized sheep. Pulmonary artery pressure, arterial P*, and rectal temperature were monitored. The media fraction, but not the bacteria, contained a component (molecular weight, 2 x lo6) composed of 84% carbohydrate and 16% protein with physiblogic~ activity. Two mg quantities, when infused, caused the ~ulmonarv artery pressure to increase 100%. Po2 to decrease by iO% and -chills -i d fever. The active component could be degraded by hot phenol-water extraction into a pure polysaccharide (molecular weight, 200,000). This lower molecular weight polysaccharide retained the identical physiologic properties when infused in the sheep. The degraded component precipitated with group B-specific antiserum.This study demonstrates that, in the sheep, a pure polysaccharide is the physiologically active part of a high-molecular-weight toxin synthesized by group B b-hemolytic Streptococcus type 111 and that this component has a different carbohydrate composition from the group ~-c a~s u l a r antigen. SpeculationThe clinical syndrome associated with group B /3-hemolytic Streptococci in early onset disease is caused by the interactions of an extracellular bacterial component and a specific target tissue in the infected infant.Group B P-hemolytic Streptococcus has become a major pathogen in newborn nurseries in the United States (19). Two distinct clinical syndromes have been described (1) depending on the age of presentation. The early onset disease, characterized by a shortlived picture and with a mortality rate in some series of over 50% (18,34), has been compared clinically by many investigators (13,20,25) to gram-negative endotoxin shock. Although the clinical and laboratory picture suggests the presence of bacterial products with endotoxin-like properties, such products have so far not been identified.The presence of extracellular toxins has been described for a number of strains of group A hemolytic Streptococci. They have been associated with the erythema of scarlet fever, a pyrogenic response in rabbits, and the enhancement of gram-negative endotoxin shock in mice, monkeys, and rabbits (10,21,38). Several workers have attempted to characterize potentially pathogenic extracellular products produced by group B P-hemolytic Streptococci. Todd, in 1934 (33), described the production of an oxygen stable, non-immunogenic hemolysin; McClean, in 1941 (26), demonstrated that these organisms elaborated a hyaluronidase; Brown et al., in 1974 (6), reported the isolation of the protein's cyclic adenosine 3'5'-monophosphate factor; and Milligan et al. in 1977 (27), described the presence of neuroaminidase (sialidase) in concentrated culture filtrates. The role of these products in the pathoph...

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Section: Control For Gram-negative Contaminationmentioning

confidence: 99%

Studies on Group B β-Hemolytic Streptococcus. I. Isolation and Partial Characterization of an Extracellular Toxin

et al. 1981

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“…Obviously peaks originating from components of 0 side chains disappear; the peaks A, D, E, G, H, and most of peak B belong to this chss (see Table 1 and Fig.2). In addition, peak I, which was previously identified as a derivative of 3,6-di-Omethyl-D-galactose [4], was absent from the chromatogram derived from the Ra lipopolysaccharide. Since we anticipated that a di-0-methyl-glucose originating from the subterminal glucose residue of R core will be found in S lipopolysaccharide but not in Ra lipopolysaccharide (see Introduction), we examined the possibility that this peak might actually be derived from a di-0-methyl-D-glucose rather than a di-0-methyl-D-galactose.…”

Section: Comparison Of S and Ra Lipopolysaccharidementioning

confidence: 83%

“…Since this peak was identified by Hellerqvist et al [3,4] as a derivative of 3,6-di-O-methyl-hexitol on the basis of mass spectrometric analysis, and since D-mannose, D-galactose, and D-glucose are the only hexoses found in S. typhimurium lipopolysaccharide, peak I is expected to be a derivative of one of the three sugars just mentioned. A uniformly 14C-labeled lipopolysaccharide was prepared with an S strain, LT2, and after methylation, hydrolysis, reduction, and acetylation, the methyl acetyl alditols were separated by gas-liquid chromatography.…”

Section: Idmtification Of Peak Imentioning

confidence: 92%

“…Presumably the stock of LT2 strain used by Hellerqvist et al [4] was predominantly in its 12,-positive form.…”

Section: Comparison Of S and Ra Lipopolysaccharidementioning

confidence: 99%

“…The methylation analysis of complex heteropolysaccharides was greatly improved by Lindberg and his coworkers, who converted methylated sugars into 0-acetyl-0-methyl-alditois before gas-liquid chromatography, and identified peaks by mass spectrometric analysis. A thorough analysis of Salmonella typhimurium cell wall lipopolysaccharide by this technique has been published [3,4], but none of the expected derivatives of subterminal glucose was seen. No tri-0-methyl-glucose was found, and although one di-0-methyl-glucose was seen, it had 0-methyl groups on C-2 and (2-4, whereas the expected di-0-methyl-glucose from the linkage region should not be methylated a t C-4 as this position would be blocked by the 0 side chain [2].…”

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confidence: 99%

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Structure of Cell Wall Lipopolysaccharide from Salmonella typhimurium. Further Studies on the Linkage between O Side Chains and R Core

Nikaido

1970

Eur J Biochem

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Cell wall lipopolysaccharide from a wild type Salmonella typhimurium and its Ra mutant were methylated and hydrolyzed, and methylated sugars were examined by gas-liquid chromatography after reduction and acetylation. A peak from the wild type lipopolysaccharide, previously believed to be the derivative of 3,6-di-O-methyl-~-galactose, was found to be derived from 3,6-di-O-methyl-~-glucose by the use of uniformly 14C-labeled lipopolysaccharide and of lipopolysaccharide specifically labeled with 14C in its galactose residues. Ra mutant lipopolysaccharide, which lacks the 0 side chain portion, did not produce the above-mentioned derivative but produced upon methylation a sugar, tentatively identified from its retention time as 3,4,6-tri-O-methyl-D-glucose. These results are consistent with our earlier proposal that 0 side chains are linked to C-4 of the subterminal D-glucose residue of R core portion of lipopolysaccharide, and further indicate that this glucose residue is also substituted a t C-2, presumably by the non-reducing terminal N-acetyl-D-glucosamine residue of the R core.Cell wall lipopolysaccharide from Salmonella consists of three regions [l] (Fig. 1). The outermost region, 0 side chain, which is composed of oligosaccharide repeat units, is linked to the R core polysaccharide, which is then linked to a lipid, called lipid A. I n a preceding paper of this series, it was shown, by periodate oxidation, that 0 side chains are However, owing to the limitations inherent in the methods used, the conclusion was not definite and confirmation by other methods was thought desirable. This paper describes the results obtained by one such method, i.e. methylation analysis.If the structure of linkage region proposed earlier is correct (see Fig. l), we can make several predictions on the outcome of methylation of cell wall lipopolysaccharide followed by acid hydrolysis. The methylation analysis of complex heteropolysaccharides was greatly improved by Lindberg and his coworkers, who converted methylated sugars into 0-acetyl-0-methyl-alditois before gas-liquid chromatography, and identified peaks by mass spectrometric analysis. A thorough analysis of Salmonella typhimurium cell wall lipopolysaccharide by this technique has been published [3,4], but none of the expected derivatives of subterminal glucose was seen. No tri-0-methyl-glucose was found, and although one di-0-methyl-glucose was seen, it had 0-methyl groups on C-2 and (2-4, whereas the expected di-0-methyl-glucose from the linkage region should not be methylated a t C-4 as this position would be blocked by the 0 side chain [2]. METHODS Bacterial StrainsSalmonella typhirnurium LT2 and its derivatives were nsed. TV205 is a rfb mutant, synthesizing an Ra-type lipopolysaccharide composed of a complete

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The Determination of Carbohydrate in Biological Materials by Gas‐Liquid Chromatography

Clamp

Bhatti

Chambers

1971

Methods of Biochemical Analysis

207

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Structural studies on the O-specific side-chains of the cell-wall lipopolysaccharide from Salmonella typhimurium LT2

Cited by 87 publications

References 5 publications

Studies on Group B β-Hemolytic Streptococcus. I. Isolation and Partial Characterization of an Extracellular Toxin

Studies on Group B β-Hemolytic Streptococcus. I. Isolation and Partial Characterization of an Extracellular Toxin

Structure of Cell Wall Lipopolysaccharide from Salmonella typhimurium. Further Studies on the Linkage between O Side Chains and R Core

The Determination of Carbohydrate in Biological Materials by Gas‐Liquid Chromatography

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