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PMID: 3872843 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Composition and antigenic activity of the oligosaccharide moiety of Haemophilus influenzae type b lipooligosaccharide.

Infection and immunity ·Vol. 48 ·No. 2 ·1985-05-00 ·Pages 324-30

Inzana TJ, Seifert WE, Williams RP

Abstract

The oligosaccharide moiety of the lipooligosaccharide of Haemophilus influenzae type b strain Eag was isolated from the lipid component by mild acid hydrolysis and purified by gel filtration. Fast atom bombardment-mass spectrometry indicated that the lipid-free oligosaccharide had a basic molecular weight of 1,768; polysaccharides comparable to high-molecular-weight O side chains were not found. Glucose, galactose, galactosamine, heptose, 3-deoxy-D-manno-2-octulosonic acid (KDO), ethanolamine, and phosphate were identified in the lipid-free oligosaccharide by colorimetric assays, gas chromatography-mass spectrometry, or an amino acid analyzer. The presence of KDO was not clearly established by a thiobarbituric acid assay or by growth inhibition by a diazaborine derivative thought to block KDO synthesis. However, the semicarbizide assay and gas chromatography-mass spectrometry confirmed the presence of KDO. Lectin precipitation by Eag lipooligosaccharide in gels indicated that beta-D-galactose was present and that some of this monosaccharide was a terminal, nonreducing residue linked to N-acetyl-D-galactosamine. The lipid-free oligosaccharide was antigenic and completely inhibited lipooligosaccharide antibody (predominantly immunoglobulin G [IgG] and IgM) in an enzyme-linked immunosorbent assay, whereas the solubilized lipid A moiety did not. H. influenzae type b lipid-free oligosaccharide differed from core oligosaccharide of Salmonella lipooligosaccharide by the presence of galactosamine and a smaller percentage of heptose and KDO.

MeSH Terms
Ethanolamines/analysis Galactosamine/analysis Galactose/analysis Glucose/analysis Haemophilus influenzae/analysis,immunology Heptoses/analysis Lipopolysaccharides/analysis,immunology Molecular Weight Oligosaccharides/analysis,immunology Phosphates/analysis Sugar Acids/analysis
Chemicals
Ethanolamines Heptoses Lipopolysaccharides Oligosaccharides Phosphates Sugar Acids 2-keto-3-deoxyoctonate Galactosamine Glucose Galactose
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Inzana T J
Seifert W E
Williams R P
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37 references, click to expand
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Article Info
Journal
Infection and immunity
Abbr.
Infect Immun
ISSN
0019-9567
DOI
10.1128/iai.48.2.324-330.1985
Published
1985-05-00
Pages
324-30
Language
English
Region
United States
NLM ID
0246127
PMCID
PMC261308
Subset
IM
Grants
NIAID NIH HHS · AI07145 · United States
NCRR NIH HHS · RR00317 · United States
External Links
PubMed source DOI: 10.1128/iai.48.2.324-330.1985 PMC Full Text View journal details
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