Genetic basis for expression of the major globotetraose containing lipopolysaccharide from Haemophilus influenzae strain Rd (RM118)

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DOIResolve DOI: http://doi.org/10.1093/glycob/11.11.957
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TypeArticle
Journal titleGlycobiology
ISSN0959-6658
1460-2423
Volume11
Issue11
Pages957967
Subjectglobotetraose; Haemophilus influenzae; lipopolysaccharide
AbstractA genetic basis for the biosynthetic assembly of the globotetraose containing lipopolysaccharide (LPS) of Haemophilus influenzae strain RM118 (Rd) was determined by structural analysis of LPS derived from mutant strains. We have previously shown that the parent strain RM118 elaborates a population of LPS molecules made up of a series of related glycoforms differing in the degree of oligosaccharide chain extension from the distal heptose residue of a conserved phosphorylated inner-core element, l-α-d-Hepp-(1→2)-L-α-d-Hepp-(1→3)-[β-d-Glcp-(1→4)-]-l-α-d-Hepp-(1→5)-α-Kdo. The fully extended LPS glycoform expresses the globotetraose structure, β-d-GalpNAc-(1→3)-α-d-Galp-(1→4)-β-d-Galp-(1→4)-β-d-Glcp. A fingerprinting strategy was employed to establish the structure of LPS from strains mutated in putative glycosyltransferase genes compared to the parent strain. This involved glycose and linkage analysis on intact LPS samples and analysis of O-deacylated LPS samples by electrospray ionization mass spectrometry and 1D 1H-nuclear magnetic resonance spectroscopy. Four genes, lpsA, lic2A, lgtC, and lgtD, were required for sequential addition of the glycoses to the terminal inner-core heptose to give the globotetraose structure. lgtC and lgtD were shown to encode glycosyltransferases by enzymatic assays with synthetic acceptor molecules. This is the first genetic blueprint determined for H. influenzae LPS oligosaccharide biosynthesis, identifying genes involved in the addition of each glycose residue.
Publication date
PublisherOxford University Press
LanguageEnglish
AffiliationNRC Institute for Biological Sciences; National Research Council Canada
Peer reviewedYes
NRC numberHOOD2001
NPARC number9358931
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Record identifier33ec888e-b4f7-43b3-ae36-fa88ebdce74e
Record created2009-07-10
Record modified2017-07-17
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