Taxonomic group: bacteria / Proteobacteria
(Phylum: Proteobacteria)
Associated disease: infection due to Escherichia coli [ICD11:
XN6P4 
]
NCBI PubMed ID: 9266718Publication DOI: 10.1111/j.1432-1033.1997.00716.xJournal NLM ID: 0107600Publisher: Oxford, UK: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
Institutions: Division of Medical and Biochemical Microbiology, Research Center Borstel, Center for Medicine and Biosciences, Borstel, Germany
The chromosomal genes rfaC and rfaF of Escherichia coli W3110 were inactivated by allelic-replacement mutagenesis to generate a defined strain lacking both heptosyltransferases which catalyze in lipopolysaccharide (LPS) biosynthesis the transfer of the first two L-glycero-D-manno-heptose (Hep) residues to 3-deoxy-D-manno-2-octulosonic acid (Kdo). The LPS of the mutant was isolated and its chemical structure was investigated by compositional analysis and nuclear magnetic resonance spectroscopy of isolated, deacylated oligosaccharide phosphates. The basic structure was a tetrasaccharide α-Kdo-(2→4)-α-Kdo-(2→6)-β-D-GlcN4P-(1→6)-α-D-GlcN1P which in LPS was substituted at position 07 of Kdo II by 2-aminoethanol phosphate in non-stoichiometric amounts. 2-Aminoethanol was cleaved during deacylation of the LPS by successive hydrazinolysis and KOH treatment and, in addition, phosphate migration from 07 to 08 of Kdo II occurred. Thus, the oligosaccharides α-Kdo7P-(2→4)-α-Kdo-(2→6)-β-D-GlcN4P-(1→6)-α-D-GlcN1P and α-Kdo8P-(2→4)-α-Kdo-(2→6)-β-D-GlcN4P-(1→6)-α-D-GlcN1P could be isolated. KOH treatment of the two trisphosphates and authentic methyl 3-deoxy-D-manno-octulopyranoside 7-(2-acetamidoethyl phosphate) proved that phosphate migration only took place when the phosphate group was substituted with 2-aminoethanol. Complementation studies with plasmid-encoded rfaC and rfaF genes revealed that the mutant strain can be used in combination with LPS-specific antibodies for the cloning and characterization of heptosytransferases which glycosylate Kdo residues of the inner core region of LPS.
Lipopolysaccharide, heptosyltransferase, phosphate migration, Escherichia coli K-12, Re mutant, rfaC
Structure type: oligomer
Location inside paper: Fig. 3, compound 1
Trivial name: core-lipid A carbohydrate backbone
Compound class: core oligosaccharide, LPS
Contained glycoepitopes: IEDB_130650,IEDB_130659,IEDB_135394,IEDB_141807,IEDB_150908,IEDB_151531
Methods: NMR, HPAEC
Biosynthesis and genetic data: genetic data
Related record ID(s): 2426, 2541, 4475, 5691, 6288, 6370, 6371, 6419, 25552, 124180
NCBI Taxonomy refs (TaxIDs): 83333Reference(s) to other database(s): GlycomeDB:
8179, CCSD:
46733, CBank-STR:13726
Show glycosyltransferases
There is only one chemically distinct structure:
Found 
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