Taxonomic group: bacteria / Proteobacteria
(Phylum: Proteobacteria)
Associated disease: infection due to Escherichia coli [ICD11:
XN6P4 
]
The structure was elucidated in this paperNCBI PubMed ID: 17316025Journal NLM ID: 0370623Publisher: American Chemical Society
Correspondence: mtanner
chem.ubc.ca
Institutions: Department of Chemistry, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z1
ADP-L-glycero-D-manno-heptose 6-epimerase (HldD or AGME, formerly RfaD) catalyzes the inversion of configuration at C-6" of the heptose moiety of ADP-D-glycero-D-manno-heptose and ADP-L-glycero-D-manno-heptose. The epimerase HldD operates in the biosynthetic pathway of L-glycero-D-manno-heptose, which is a conserved sugar in the core region of lipopolysaccharide (LPS) of Gram-negative bacteria. Previous studies support a mechanism in which HldD uses its tightly bound NADP+ cofactor to oxidize directly at C-6", generating a ketone intermediate. A reduction of the ketone from the opposite face then occurs, generating the epimeric product. How the epimerase is able access both faces of the ketone intermediate with correct alignment of the three required components, NADPH, the ketone carbonyl, and a catalytic acid/base residue, is addressed here. It is proposed that the epimerase active site contains two catalytic pockets, each of which bears a catalytic acid/base residue that facilitates reduction of the C-6" ketone but leads to a distinct epimeric product. The ketone carbonyl may access either pocket via rotation about the C-5"-C-6" bond of the sugar nucleotide and in doing so presents opposing faces to the bound cofactor. Evidence in support of the two-base mechanism is found in studies of two single mutants of the Escherichia coli K-12 epimerase, Y140F and K178M, both of which have severely compromised epimerase activities that are more than 3 orders of magnitude lower than that of the wild type. The catalytic competency of these two mutants in promoting redox chemistry is demonstrated with an alternate catalytic activity that requires only one catalytic base: dismutation of a C-6" aldehyde substrate analogue (ADP-β-D-manno-hexodialdose) to an acid and an alcohol (ADP-β-d-mannuronic acid and ADP-β-D-mannose). This study identifies the two catalytic bases as tyrosine 140 and lysine 178. A one-step enzymatic conversion of mannose into ADP-β-mannose is also described and used to make C-6"-substituted derivatives of this sugar nucleotide
Lipopolysaccharide, biosynthesis, L-glycero-D-manno-heptose, epimerase, sugar nucleotide, Escherichia coli K12
Structure type: monomer
Location inside paper: p.3917, fig,1
Trivial name: ADP-β-D-glycero-D-manno-heptose
Contained glycoepitopes: IEDB_137353,IEDB_140947
Methods: 1H NMR, 31P NMR, ESI-MS, chemical methods, genetic methods
Enzymes that release or process the structure: ADP-L-glycero-D-manno-heptose 6-epimerase (HldD)
Biosynthesis and genetic data: genetic data, enzymatic data
Synthetic data: chemical and enzymatic
Related record ID(s): 941, 944, 21501, 21868
NCBI Taxonomy refs (TaxIDs): 316407Reference(s) to other database(s): GlycomeDB:
25347
Show glycosyltransferases
There is only one chemically distinct structure:
Found 
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