Structural studies of the O-specific polysaccharide of Vibriocholerae O8 using solvolysis with triflic acid

Kocharova, Nina A.; Perepelov, Andrei V.; Zatonsky, George V.; Shashkov, A. S.; Knirel, Yuriy A.; Jansson, Per‐Erik; Weintraub, Andrej

doi:10.1016/s0008-6215(00)00271-8

Cited by 24 publications

(16 citation statements)

References 20 publications

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“…The strains were collected from patients admitted with a diagnosis of diarrhea to the Infectious Disease Hospital by the National Institute of Enteric Disease in Kolkata (formerly known as Calcutta), India, in 1994 and 1995. These isolates were collected as part of routine surveillance (26,34). The strains were selected based on overlapping sugar composition (see Introduction) and ease of long-range PCR amplification.…”

Section: Methodsmentioning

confidence: 99%

“…Vibrio cholerae O antigens include a number of sugars that are somewhat unusual, including quinovosamine (O139 and O108) and fucosamine. 2,4-Diacet-amido-2,4,6-trideoxyglucose (QuiNAc4NAc) has been reported in both O8 (26) and O5 serogroups (22). In addition, glycero-D-manno-heptose (O5, O8, and O108), glucose (O5 and O8), fructose (O5 and O8), glucosamine (O108), galactosamine (O108), and fucosamine (O108) had been reported in these serogroups (28).…”

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confidence: 99%

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Genetic Diversity of O-Antigen Biosynthesis Regions in Vibrio cholerae

Aydanian

Tang

Morris

et al. 2011

Appl Environ Microbiol

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O-antigen biosynthetic (wbf) regions for Vibrio cholerae serogroups O5, O8, and O108 were isolated and sequenced. Sequences were compared to those of other published V. cholerae O-antigen regions. These wbf regions showed a high degree of heterogeneity both in gene content and in gene order. Genes identified frequently showed greater similarities to polysaccharide biosynthesis genes from species other than V. cholerae. Our results demonstrate the plasticity of O-antigen genes in V. cholerae, the diversity of the genetic pool from which they are drawn, and the likelihood that new pandemic serogroups will emerge.Cholera is a pandemic diarrheal disease that continues to be an important cause of morbidity and mortality worldwide. Cholera is associated with a clonally related subset of Vibrio cholerae strains, which carry ctxAB (cholera toxin subunits A and B), the vibrio pathogenicity island (VPI), and other cholera-associated genes (15,40). Until 1992, only serogroup O1 (out of more than 206 serogroups currently described) was recognized as a cause of cholera. In 1992, a new, non-O1 V. cholerae strain (subsequently designated V. cholerae O139) appeared in India and rapidly spread across much of Asia (32). Extensive studies (5,12,31,41) demonstrated that the O139 strain was closely related to the O1 El Tor strains of the 7th pandemic, except that the genes responsible for O1-antigen biosynthesis were deleted and replaced with DNA that encodes the O139 antigen. Since the O antigen is the major protective epitope, its alteration was sufficient to allow O139 strains to move in epidemic form through populations previously immune to cholera caused by O1. Thus, adults were more commonly affected than children (1). Recent studies suggest that the pandemic cluster carries other serogroups as well, including O37, O27, O53, O65, and O75 (31, 40, 43). These observations are consistent with the hypothesis that pathogenic V. cholerae strains are able to easily acquire and/or exchange O-antigen genes, with the new O antigen allowing strains to evade preexisting immunity to cholera.Changes in O-antigen structure also may provide selective advantages in the environment. During epidemics, bacteriophage may play a crucial role in controlling the number of V. cholerae in the environment (19), and since the O-antigen may serve as bacteriophage receptors, serogroup conversion also may be beneficial for evading phage predation (33). It is well documented that mobile genetic elements, bacteriophage, and the competence of V. cholerae to take up and assimilate free DNA from the environment significantly contribute to genetic diversity in V. cholerae (18). Serogroup conversion was demonstrated while V. cholerae was growing on a chitin substrate (6). This transfer may have been facilitated by the JUMPstart sequence, which has nucleotide similarity to a DNA uptake signal that causes the preferential uptake of free DNA containing that sequence in the Pasteurellaceae and is present in front of a laterally transferred O-antigen region in V. chole...

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Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

Genetic Diversity of O-Antigen Biosynthesis Regions in Vibrio cholerae

Aydanian

Tang

Morris

et al. 2011

Appl Environ Microbiol

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“…, II)-By comparing chemical shifts (␦ ppm) and coupling constants (J H,H ) to those reported in the literature (35,36), the UDP-diacetamido-trideoxyhexose metabolite (II) was identified as UDP-2,4-diacetamido-2,4,6-trideoxy-␣-D-glucopyranose (UDP-6-deoxy-␣-D-GlcNAc4NAc). The 1 H NMR spectrum acquired at 600 MHz ( 1 H) with the cold probe revealed sharp peaks characteristic of UDP-6-deoxy-␣-D-GlcNAc4NAc (Fig.…”

Section: Udp-24-diacetamido-246-trideoxy-␣-d-glucopyranose (Udp-6-mentioning

confidence: 96%

Functional Characterization of the Flagellar Glycosylation Locus in Campylobacter jejuni 81–176 Using a Focused Metabolomics Approach

McNally

Hui

Aubry

et al. 2006

Journal of Biological Chemistry

110

129

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Bacterial genome sequencing has provided a wealth of genetic data. However, the definitive functional characterization of hypothetical open reading frames and novel biosynthetic genes remains challenging. This is particularly true for genes involved in protein glycosylation because the isolation of their glycan moieties is often problematic. We have developed a focused metabolomics approach to define the function of flagellin glycosylation genes in Campylobacter jejuni 81-176. A capillary electrophoresis-electrospray mass spectrometry and precursor ion scanning method was used to examine cell lysates of C. jejuni 81-176 for sugar nucleotides. Novel nucleotide-activated intermediates of the pseudaminic acid (Pse5NAc7NAc) pathway and its acetamidino derivative (PseAm) were found to accumulate within select isogenic mutants, and use of a hydrophilic interaction liquid chromatography-mass spectrometry method permitted large scale purifications of the intermediates. NMR with cryo probe (cold probe) technology was utilized to complete the structural characterization of microgram quantities of CMP-5-acetamido-7-acetamidino-3,5,7,9-tetradeoxy-L-glycero-␣-L-manno-nonulosonic acid (CMP-Pse5NAc7Am), which is the first report of Pse modified at C7 with an acetamidino group in Campylobacter, and UDP-2,4-diacetamido-2,4,6-trideoxy-␣-D-glucopyranose, which is a bacillosamine derivative found in the N-linkedproteinglycan.Usingthisfocusedmetabolomicsapproach,pseB, pseC, pseF, pseI, and for the first time pseA, pseG, and pseH were found to be directly involved in either the biosynthesis of CMP-Pse5NAc7NAc or CMP-Pse5NAc7Am. In contrast, it was shown that pseD, pseE, Cj1314c, Cj1315c, Cjb1301, Cj1334, Cj1341c, and Cj1342c have no role in the CM-P-Pse5NAc7NAc or CMP-Pse5NAc7Am pathways. These results demonstrate the usefulness of this approach for targeting compounds within the bacterial metabolome to assign function to genes, identify metabolic intermediates, and elucidate novel biosynthetic pathways.The availability of bacterial genomic sequences has provided unprecedented opportunity for comparative studies. The functional analysis of each respective genome is currently under way and often involves an integrative, multidisciplinary approach that combines bioinformatics, mutagenesis, proteomics, and microarray technologies. Most recently, metabolomics-based analyses, although limited in their number, are becoming established as an additional tool to elucidate gene function (1, 2). Metabolomics is the characterization of all low molecular weight compounds in a defined biological system and differs from classical metabolism studies by its greater breadth and speed of metabolite analysis. Although recent advances in mass spectrometry (MS), 2 such as the ultimate resolving capability of the Fourier-transform ion cyclotron resonance MS, and also in nuclear magnetic resonance spectroscopy (NMR) technologies, such as the development of higher magnetic field spectrometers and cryogenically cooled probes (cold probes), has greatly facilit...

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“…One example is the use of NMR and MS in combination for the structural determination of the O-specific polysaccharide of Vibrio cholerae O8. 4 NMR studies can also provide information about the stereochemical configuration, especially the anomeric configuration, of the monomers in the polymer chains. 5 In general, when obtaining spectra from higher molecular weight oligosaccharides and polymers, the characteristic signals from the end residues diminishes and finally disappears when the molecular weight exceeds a certain limit.…”

Section: Determination Of Monosaccharide Compositionmentioning

confidence: 99%