Electrophoretic and Mass Spectrometric Strategies for the Identification of Lipopolysaccharides and Immunodeterminants in Pathogenic Strains of Haemophilus influenzae; Application to Clinical Isolates

Thibault, Pierre; Li, J.; Martin, Andrew; Richards, James C.; Hood, Derek W.; Moxon, E. Richard

doi:10.1007/978-1-59259-719-2_22

Cited by 9 publications

(4 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The fragmentation of cationic oligosaccharides typically proceeds by cleavage at the glycosidic bonds, which provides sequence and branching information (52). The product ion spectrum (MS/MS spectrum) obtained from doubly charged ion at m/z 1255.12 is illustrated in Fig.…”

Section: Da) Sequence Information Was Further Characterized By Tandementioning

confidence: 99%

Chromosomal and Plasmid-encoded Enzymes Are Required for Assembly of the R3-type Core Oligosaccharide in the Lipopolysaccharide of Escherichia coli O157:H7

Kaniuk

Vinogradov

et al. 2004

Journal of Biological Chemistry

View full text Add to dashboard Cite

The type R3 core oligosaccharide predominates in the lipopolysaccharides from enterohemorrhagic Escherichia coli isolates including O157:H7. The R3 core biosynthesis (waa) genetic locus contains two genes, waaD and waaJ, that are predicted to encode glycosyltransferases involved in completion of the outer core. Through determination of the structures of the lipopolysaccharide core in precise mutants and biochemical analyses of enzyme activities, WaaJ was shown to be a UDP-glucose:(galactosyl) lipopolysaccharide ␣-1,2-glucosyltransferase, and WaaD was shown to be a UDP-glucose:(glucosyl)lipopolysaccharide ␣-1,2-glucosyltransferase. The residue added by WaaJ was identified as the ligation site for O polysaccharide, and this was confirmed by determination of the structure of the linkage region in serotype O157 lipopolysaccharide. The initial O157 repeat unit begins with an N-acetylgalactosamine residue in a ␤-anomeric configuration, whereas the biological repeat unit for O157 contains ␣-linked N-acetylgalactosamine residues. With the characterization of WaaJ and WaaD, the activities of all of the enzymes encoded by the R3 waa locus are either known or predicted from homology data with a high level of confidence. However, when core oligosaccharide structure is considered, the origin of an additional ␣-1,3-linked N-acetylglucosamine residue in the outer core is unknown. The gene responsible for a nonstoichiometric ␣-1,7-linked N-acetylglucosamine substituent in the heptose (inner core) region was identified on the large virulence plasmids of E. coli O157 and Shigella flexneri serotype 2a. This is the first plasmid-encoded core oligosaccharide biosynthesis enzyme reported in E. coli.

show abstract

Section: Da) Sequence Information Was Further Characterized By Tandementioning

confidence: 99%

Chromosomal and Plasmid-encoded Enzymes Are Required for Assembly of the R3-type Core Oligosaccharide in the Lipopolysaccharide of Escherichia coli O157:H7

Kaniuk

Vinogradov

et al. 2004

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…During the last few years our laboratory has developed on-line CE-MS techniques for trace level analysis of LPS glycoform and isoform distributions from a range of bacterial species. [16][17][18][19][20] In this article we report on the application CE-MS to the profiling of isomeric glycoforms and phosphoforms from H. influenzae LPS. The exceptional analytical potential of CE-MS to separate closely related glycoform families is *Jianjun.Li@nrc-cnrc.gc.ca demonstrated for LPS samples obtained from two H. influenzae strains, Rd11.7 and 375.1, that differ in glycoform composition and phosphoethanolamine substitution patterns.…”

Section: Introductionmentioning

confidence: 99%

Electrophoretic and mass spectrometric strategies for profiling bacterial lipopolysaccharides

Cox

Hood

et al. 2005

Mol. Biosyst.

View full text Add to dashboard Cite

Capillary electrophoresis (CE) is a high-resolution separation technique that has been widely used for trace analysis in biological samples. On-line capillary electrophoresis-electrospray mass spectrometry (CE-MS) was developed for the analysis of lipopolysaccharide (LPS) glycoforms from the gram-negative bacteria, Haemophilus influenzae. In this paper, we report on the application of CE-MS to characterize structural differences in O-deacylated LPS samples from H. influenzae strains Rd 11.7 and 375.1. The resolution capability of on-line CE-MS was first demonstrated by analysis of a complex LPS mixture from H. influenzae strain Rd 11.7. This strain contains a mixture of isomeric glycoforms differing in the number and positions of hexose moieties. Sialic acid containing glycoforms were also determined. Structural features of LPS from a lic1 mutant of H. influenzae strain 375 (375.1) were studied using on-line CE-MS/MS. With the separation provided by CE, two isomeric glycoforms differing in the location of phosphoethanolamine substituents were characterized by tandem mass spectrometry.

show abstract

“…The composition details, as well as some sequence information of those two major components with m / z 1180.24 and 1197.25, were further characterized by tandem mass spectrometry (MS/MS). The fragmentation of cationic oligosaccharides typically proceeds by cleavage at the glycosidic bonds, which provides sequence and branching information [31]. The charge state of a fragment ion is then identified by using the isotope profile, owing to the high resolution provided by the TOF mass analyser.…”

Section: Resultsmentioning

confidence: 99%

The structure and biological characteristics of the Spirochaeta aurantia outer membrane glycolipid LGL_B

Vinogradov

Paul

et al. 2004

European Journal of Biochemistry

View full text Add to dashboard Cite

In an attempt to isolate lipopolysaccharide from Spirochaeta aurantia, Darveau-Hancock extraction of the cell mass was performed. While no lipopolysaccharide was found, two carbohydrate-containing compounds were detected. They were resolved by size-exclusion chromatography into high molecular mass (LGL A ) and low molecular mass (LGL B ) fractions. Here we present the results of the analysis of the glycolipid LGL B . Deacylation of LGL B with hydrazine and separation of the products by using anion-exchange chromatography gave two major products. Their structure was determined by using chemical methods, NMR and mass spectrometry. All monosaccharides had the D-configuration, and aspartic acid had the L-configuration. Intact LGL B contained two fatty groups at O-2 and O-3 of the glycerol residue. Nonhydroxylated C14 to C18 fatty acids were identified, which were predominantly unsaturated or branched.LGL B was able to gel Limulus amebocyte lysate, albeit at a lower level than that observed for Escherichia coli O113 lipopolysaccharide. However, even large amounts of LGL B were unable to stimulate any Toll-like receptor (TLR) examined, including TLR4 and TLR2, previously shown to be sensitive to lipopolysaccharide and glycolipids from diverse bacterial origins, including other spirochetes.

show abstract

Electrophoretic and Mass Spectrometric Strategies for the Identification of Lipopolysaccharides and Immunodeterminants in Pathogenic Strains of Haemophilus influenzae; Application to Clinical Isolates

Cited by 9 publications

References 31 publications

Chromosomal and Plasmid-encoded Enzymes Are Required for Assembly of the R3-type Core Oligosaccharide in the Lipopolysaccharide of Escherichia coli O157:H7

Chromosomal and Plasmid-encoded Enzymes Are Required for Assembly of the R3-type Core Oligosaccharide in the Lipopolysaccharide of Escherichia coli O157:H7

Electrophoretic and mass spectrometric strategies for profiling bacterial lipopolysaccharides

The structure and biological characteristics of the Spirochaeta aurantia outer membrane glycolipid LGL_B

Contact Info

Product

Resources

About

Electrophoretic and Mass Spectrometric Strategies for the Identification of Lipopolysaccharides and Immunodeterminants in Pathogenic Strains of Haemophilus influenzae; Application to Clinical Isolates

Cited by 9 publications

References 31 publications

Chromosomal and Plasmid-encoded Enzymes Are Required for Assembly of the R3-type Core Oligosaccharide in the Lipopolysaccharide of Escherichia coli O157:H7

Chromosomal and Plasmid-encoded Enzymes Are Required for Assembly of the R3-type Core Oligosaccharide in the Lipopolysaccharide of Escherichia coli O157:H7

Electrophoretic and mass spectrometric strategies for profiling bacterial lipopolysaccharides

The structure and biological characteristics of the Spirochaeta aurantia outer membrane glycolipid LGLB

Contact Info

Product

Resources

About

The structure and biological characteristics of the Spirochaeta aurantia outer membrane glycolipid LGL_B