Structures of lipopolysaccharides from Klebsiella pneumoniae - Elucidation of the structure of the linkage region between core and polysaccharide O chain and identification of the residues at the non-reducing termini of the O chains

E. Vinogradov, E. Frirdich, L. L. MacLean, M. B. Perry, Bent O. Petersen, Jens Øllgaard Duus, C. Whitfield

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Deamination of LPSs from Klebsiella pneumoniae released O-chain polysaccharides together with a fragment of the core oligosaccharide. The structures of the products from serotypes 01, O2a, O2a,c, 03, 04, 05, and 012 were determined by NMR spectroscopy and chemical methods, identifying the linkage region between the 0 antigens and the core as well as novel residues at the non-reducing ends of the polysaccharides. All serotypes had an identical linkage between the 0 chain and core. [GRAPHICS] where L-glycero-D-manno-heptose is non-stoichiometric. Analysis of the LPSs from a waaL (O-polysaccharide ligase-deficient) mutant showed that the beta-GlcNAc in the linkage region is derived from the O-chain biosynthesis pathway, and the alpha-3-deoxy-D-manno-octulosonic acid represents the core residue to which the 0 chain is ligated. Consistent with the common ligation site, the WaaL proteins from these serotypes are essentially identical. Polysaccharides from serotypes 01, O2a, and O2a,c contain no special groups at the non-reducing end, whereas polysaccharides from serotypes 03,04,05, and 012 have residues at the non-reducing end that are not found in the polymer repeating units.
Original languageEnglish
JournalJournal of Biological Chemistry
Volume277
Issue number28
Pages (from-to)25070-25081
Number of pages12
ISSN0021-9258
DOIs
Publication statusPublished - 2002
Externally publishedYes

Keywords

  • Carbohydrate Conformation
  • Carbohydrate Sequence
  • Klebsiella pneumoniae
  • Lipopolysaccharides
  • Molecular Sequence Data
  • Nuclear Magnetic Resonance, Biomolecular
  • Antigens
  • Biosynthesis
  • Nuclear magnetic resonance spectroscopy
  • Polymers
  • Polysaccharides
  • Biochemistry
  • bacterial protein
  • bacterium lipopolysaccharide
  • heptose
  • polymer
  • polysaccharide
  • article
  • biosynthesis
  • carbohydrate analysis
  • chemical composition
  • deamination
  • molecular interaction
  • nonhuman
  • nuclear magnetic resonance spectroscopy
  • nucleotide sequence
  • priority journal
  • serotype
  • structure analysis
  • Bacteria (microorganisms)
  • Klebsiella
  • Negibacteria
  • Ligation sites
  • T
  • X
  • BIOCHEMISTRY
  • VIBRIO-CHOLERAE O1
  • D-GALACTAN-I
  • CASSETTE TRANSPORT-SYSTEM
  • RFB GENE-CLUSTER
  • ESCHERICHIA-COLI
  • NCTC-5055 LIPOPOLYSACCHARIDE
  • SEROTYPE O1-K20
  • ANTIGEN
  • BIOSYNTHESIS
  • EXPRESSION
  • Facultatively Anaerobic Gram-Negative Rods Eubacteria Bacteria Microorganisms (Bacteria, Eubacteria, Microorganisms) - Enterobacteriaceae [06702] Klebsiella pneumoniae serovar-O1, serovar-O12, serovar-O2a, serovar-O2ac, serovar-O4, serovar-O5
  • Klebsiella pneumoniae waaL gene [Enterobacteriaceae] mutation
  • alpha-3-deoxy-D-manno-octulosonic acid
  • antigen
  • lipopolysaccharide deamination
  • O-polysaccharide ligase
  • WaaL protein
  • 03502, Genetics - General
  • 10062, Biochemistry studies - Nucleic acids, purines and pyrimidines
  • 10064, Biochemistry studies - Proteins, peptides and amino acids
  • 10066, Biochemistry studies - Lipids
  • 10068, Biochemistry studies - Carbohydrates
  • 31000, Physiology and biochemistry of bacteria
  • 31500, Genetics of bacteria and viruses
  • Biochemistry and Molecular Biophysics
  • 7-Tesla super-conducting magnet Bruker Daltonics, laboratory equipment
  • lipopolysaccharide residue identification Molecular Biology Techniques and Chemical Characterization, identification method
  • NMR spectroscopy nuclear magnetic resonance spectroscopy Spectrum Analysis Techniques, imaging method
  • SDS-PAGE gels laboratory equipment
  • SDS-PAGE SDS-polyacrylamide gel electrophoresis Electrophoretic Techniques, detection method
  • Western immunoblotting Immunologic Techniques, identification method
  • AF146532 Genbank, EMBL, DDBJ 225886-79-3 amino acid sequence, nucleotide sequence
  • AF482003 Genbank, EMBL, DDBJ amino acid sequence, nucleotide sequence
  • AF482004 Genbank, EMBL, DDBJ amino acid sequence, nucleotide sequence
  • AF482005 Genbank, EMBL, DDBJ amino acid sequence, nucleotide sequence
  • AF482006 Genbank, EMBL, DDBJ amino acid sequence, nucleotide sequence
  • AF482007 Genbank, EMBL, DDBJ amino acid sequence, nucleotide sequence
  • Methods and Techniques
  • Molecular Genetics

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