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Literature summary for 5.1.3.20 extracted from

  • Shaik, M.M.; Zanotti, G.; Cendron, L.
    The crystal structure of ADP-L-glycero-D-manno-heptose-6-epimerase (HP0859) from Helicobacter pylori (2011), Biochim. Biophys. Acta, 1814, 1641-1647.
    View publication on PubMed
Search for more literature for 5.1.3.20

Cloned(Commentary)

Cloned (Comment) Organism
gene HP0859, expression of C-terminally His-tagged enzyme in Escherichia coli strain BL21(DE3) Helicobacter pylori

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant His-tagged enzyme, vapor diffusion technique, mixing of 20 mg/ml protein in 30 mM Tris, pH 7.5, 150 mM NaCl, with precipitant solution containing 0.2 M ammonium sulfate, 0.1 M tri sodium citrate, pH 5.6, 15% w/v PEG 4000, and 5% glycerol, 20°C, X-ray diffraction structure determination and analysis at 2.55 A resolution Helicobacter pylori

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ADP-D-glycero-D-manno-heptose Helicobacter pylori
-
 ADP-L-glycero-D-manno-heptose
-
 ?
ADP-D-glycero-D-manno-heptose Helicobacter pylori G27
-
 ADP-L-glycero-D-manno-heptose
-
 ?

Organism

Organism UniProt Comment Textmining
Helicobacter pylori B5Z7L9 gene HP0859 or rfaD
-
Helicobacter pylori G27 B5Z7L9 gene HP0859 or rfaD
-

Purification (Commentary)

Purification (Comment) Organism
recombinant C-terminally His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gell filtration Helicobacter pylori

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ADP-D-glycero-D-manno-heptose
-
 Helicobacter pylori ADP-L-glycero-D-manno-heptose
-
 ?
ADP-D-glycero-D-manno-heptose
-
 Helicobacter pylori G27 ADP-L-glycero-D-manno-heptose
-
 ?

Subunits

Subunits Comment Organism
pentamer structure modeling of the pentameric enzyme including five protein monomers and 5 NAD+ molecules, overview Helicobacter pylori

Synonyms

Synonyms Comment Organism
ADP-L-glycero-D-manno-heptose-6-epimerase
-
 Helicobacter pylori
AGME
-
 Helicobacter pylori
HP0859
-
 Helicobacter pylori

Cofactor

Cofactor Comment Organism Structure
additional information preference of the enzyme for NAD+ compared to NADP+. When NADP is bound, the binding region assumes a different conformation Helicobacter pylori
NAD+ preferred cofactor compared to NADP+, binds at the enzyme active site, binding site structure, overview Helicobacter pylori
NADP+ NAD+ is the preferred cofactor Helicobacter pylori

pI Value

Organism Comment pI Value Maximum pI Value
Helicobacter pylori sequence calculation
-
 6.8

General Information

General Information Comment Organism
malfunction a HP0859 knockout mutant shows a severe loss of lipopolysaccharide structure and a significant reduction of adhesion levels in an infection model to human stomach gastric adenocarcinoma AGS cells, if compared with the wild-type strain Helicobacter pylori
metabolism last enzyme in the pathway that produces L-glycero-D-manno-heptose starting from sedoheptulose-7-phosphate Helicobacter pylori
additional information an N-terminal seven-stranded modified Rossmann fold where the NAD+ cofactor is bound and a smaller C-terminal alpha/beta domain are responsible for the binding of the substrate Helicobacter pylori