Structural elucidation of the major Hex4 lipopolysaccharide glycoform from the lgtC mutant of Haemophilus influenzae strain Eagan: Carbohydr.Res.

  1. Get@NRC: Structural elucidation of the major Hex4 lipopolysaccharide glycoform from the lgtC mutant of Haemophilus influenzae strain Eagan: Carbohydr.Res. (Opens in a new window)
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Journal titleCarbohydr.Res.
Pages14241434; # of pages: 11
Subject31P; bacterial; Bacterial Proteins; biosynthesis; Canada; Carbohydrate Sequence; Carbon; Carbon Isotopes; chemistry; core oligosaccharide; Deuterium; DISACCHARIDE; Electrospray; ELUCIDATION; epitope; Epitopes; Esi Ms; Galactosyltransferases; GENE; genetics; Haemophilus; Haemophilus influenzae; HAEMOPHILUS-INFLUENZAE; Hexoses; IDENTIFICATION; isolation & purification; Isotopes; KDO; LIPOPOLYSACCHARIDE; Lipopolysaccharides; LOCATION; LPS; Magnetic Resonance Spectroscopy; mass spectrometry; MASS-SPECTROMETRY; metabolism; Methylation; Molecular Sequence Data; Molecular Structure; MUTANT; Mutation; NMR; oligosaccharide; Oligosaccharide Epitopes; phosphate; protein; Proteins; SAMPLES; Spectrometry,Mass,Electrospray Ionization; STRAIN; STRUCTURAL; structural elucidation; structure; TRISACCHARIDE; UNIT; Virulence; Virulence Factors
AbstractLipopolysaccharide (LPS) oligosaccharide epitopes are major virulence factors of Haemophilus influenzae. The structure of LPS glycoforms of H. influenzae type b strain Eagan containing a mutation in the gene lgtC is investigated. LgtC is involved in the biosynthesis of globoside trisaccharide [alpha-D-Galp-(1-->4)-beta-d-Galp-(1-->4)-beta-D-Glcp-(1-->], an LPS epitope implicated in the virulence of this organism. Glycose and methylation analyses provided information on the composition while electrospray ionization mass spectrometry (ESI-MS) on O-deacylated LPS (LPS-OH) indicated the major glycoform to contain 4 hexoses attached to the common H. influenzae triheptosyl inner-core unit. The structure of the Hex4 glycoform in LPS-OH and core oligosaccharide samples was determined by NMR. It consists of an l-alpha-D-HepIIIp-(1-->2)-[PEtn-->6]-l-alpha-D-HepIIp-(1-->3)-l-alpha-D-He pIp-(1-->5)-[P-->4]-alpha-D-Kdop-(2--> to which a beta-D-Glcp-(1-->4)-alpha-D-Glcp disaccharide unit is extended from HepII at the C-3 position, while HepI and HepIII are substituted at the C-4 and C-2 positions with beta-D-Glcp and beta-D-Galp, respectively. This structure corresponds to that expressed as a subpopulation in the parent strain. 31P NMR studies permitted the identification of subpopulations of LPS containing Kdo substituted at the C-4 position with monophosphate or pyrophosphoethanolamine (PPEtn). HepIII was found to be substituted with either phosphate at the C-4 position or acetate at the C-3 position, but not both of them together in the same subpopulation. The subpopulations containing phosphate and acetate at HepIII and their location have not previously been reported
Publication date
AffiliationNRC Institute for Biological Sciences; National Research Council Canada
Peer reviewedNo
NRC numberMASOUD2008A
NPARC number9372889
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Record identifiere9b57a72-d86c-44ba-abfa-af921896fc6d
Record created2009-07-10
Record modified2016-05-09
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