Structural studies utilizing 13C-enrichment of the O-antigen polysaccharide from the enterotoxigenic Escherichia coli O159 cross-reacting with Shigella dysenteriae type 4.

The structure of the O-antigen polysaccharide from Escherichia coli O159 has been determined using primarily NMR spectroscopy of the 13C-enriched polysaccharide. The sequence of the sugar residues could be determined by heteronuclear multiple bond connectivity NMR experiments. The polysaccharide is composed of a pentasaccharide repeating unit with the following structure: [sequence: see text] Matrix assisted laser desorption ionization mass spectrometry was performed on intact lipopolysaccharide and from the resulting molecular mass the O-antigen part was estimated to contain approximately 23 repeating units. Cross-reactivity of this O-antigen to that of Shigella dysenteriae type 4 was confirmed using enzyme-linked immunoabsorbant assay.

[1]  B. Rowe,et al.  For new Escherichia coli o antigens, o158, o159, o160 and o161, and two new h antigens, h53 and h54. , 2009, Acta Pathologica Microbiologica Scandinavica Section B Microbiology.

[2]  P. Jansson,et al.  Structural studies on the hexose region of the core in lipopolysaccharides from Enterobacteriaceae. , 2005, European journal of biochemistry.

[3]  A. Weintraub,et al.  Structural determination and biosynthetic studies of the O-antigenic polysaccharide from the enterohemorrhagic Escherichia coli O91 using 13C-enrichment and NMR spectroscopy. , 1999, Biochemistry.

[4]  Göran Widmalm,et al.  NMR spectroscopy of 13C‐enriched polysaccharides: application of 13C–13C TOCSY to sugars of different configuration , 1998 .

[5]  M. Wolf,et al.  Occurrence, distribution, and associations of O and H serogroups, colonization factor antigens, and toxins of enterotoxigenic Escherichia coli , 1997, Clinical microbiology reviews.

[6]  T. Cheasty,et al.  Isolation of enterotoxigenic Escherichia coli from British troops in Saudi Arabia , 1995, Epidemiology and Infection.

[7]  Wolfgang Bermel,et al.  Gradient selection in inverse heteronuclear correlation spectroscopy , 1993 .

[8]  B. Spangler,et al.  Structure and function of cholera toxin and the related Escherichia coli heat-labile enterotoxin. , 1992, Microbiological reviews.

[9]  G. Marius Clore,et al.  1H1H correlation via isotropic mixing of 13C magnetization, a new three-dimensional approach for assigning 1H and 13C spectra of 13C-enriched proteins , 1990 .

[10]  J. Sanders,et al.  Biosynthetic studies using carbon-13-COSY: the Klebsiella K3 serotype polysaccharide , 1989 .

[11]  P. Jansson,et al.  Computer-assisted structural analysis of polysaccharides with an extended version of CASPER using 1H- and 13C-n.m.r. data. , 1989, Carbohydrate research.

[12]  P. Albersheim,et al.  Determination, by methylation analysis, of the glycosyl-linkage compositions of microgram quantities of complex carbohydrates , 1983 .

[13]  J. Vliegenthart,et al.  Determination of the absolute configuration of mono-saccharides in complex carbohydrates by capillary G.L.C. , 1979, Carbohydrate research.

[14]  B. Lindberg,et al.  Assignment of absolute configuration of sugars by g.l.c. of their acetylated glycosides formed from chiral alcohols , 1978 .

[15]  N. K. Kochetkov,et al.  Antigenic polysaccharides of bacteria IV. Structure of the repeating unit of the polysaccharide chain of Shigella dysenteriae type 3 lipopolysaccharide , 1977 .

[16]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[17]  A. Jeanes,et al.  Quantitative Determination of Monosaccharides as Their Alditol Acetates by Gas Liquid Chromatography. , 1965 .

[18]  Oliver H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.

[19]  J. Sanders,et al.  Assignment of the 13C N.M.R. spectrum of the Klebsiella K3 serotype polysaccharide by COSY spectroscopy , 1989 .

[20]  C. Frasch,et al.  A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. , 1982, Analytical biochemistry.

[21]  A. Weintraub,et al.  The humoral antibody response to Bacteroides fragilis infections in humans. , 1979, Scandinavian journal of infectious diseases. Supplementum.

[22]  B. Rowe,et al.  A NEW Escherichia coli O-group, O159, associated with outbreaks of enteritis in infants. , 1976, Scandinavian journal of infectious diseases.

[23]  P. Jansson A practical guide to the methylation analysis of carbohydrates , 1976 .

[24]  W. Ewing,et al.  Identification of Enterobacteriaceae. , 1972 .