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Literature summary extracted from

  • Kim, M.; Lim, A.; Yang, S.; Park, J.; Lee, D.; Shin, D.
    Structure and in silico substrate-binding mode of ADP-L-glycero-D-manno-heptose 6-epimerase from Burkholderia thailandensis (2013), Acta Crystallogr. Sect. D, 69, 658-668.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
5.1.3.20 gene rfaD, sequence compariosns, expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3) Burkholderia thailandensis

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
5.1.3.20 purified recombinant His-tagged enzyme, hanging drop vapour diffusion method, mixing of 200 nl of 8.8 mg/ml protein in 0.43 M NaCl, 20 mM Tris-HCl, pH 8.0, with 200 nl of reservoir solution containing 35% PEG 200, 0.1 M bis-tris-HCl, pH 5.5, and equilibration against 0.05 ml of reservori solution, method optimization, X-ray diffraction structure determination and analysis at 2.6 A resolution, molecular replacement Burkholderia thailandensis

Protein Variants

EC Number Protein Variants Comment Organism
5.1.3.20 E210G site-directed mutagenesis, structure compared to the wild-type enzyme Burkholderia thailandensis
5.1.3.20 K178M site-directed mutagenesis, structure compared to the wild-type enzyme Burkholderia thailandensis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
5.1.3.20 38000
-
 x * 38000, recombinant His6-tagged enzyme, SDS-PAGE Burkholderia thailandensis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
5.1.3.20 ADP-D-glycero-D-manno-heptose Burkholderia thailandensis
-
 ADP-L-glycero-D-manno-heptose
-
 r
5.1.3.20 ADP-D-glycero-D-manno-heptose Burkholderia thailandensis E444
-
 ADP-L-glycero-D-manno-heptose
-
 r

Organism

EC Number Organism UniProt Comment Textmining
5.1.3.20 Burkholderia thailandensis Q2SY18 gene rfaD
-
5.1.3.20 Burkholderia thailandensis E444 Q2SY18 gene rfaD
-

Purification (Commentary)

EC Number Purification (Comment) Organism
5.1.3.20 recombinant N-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity and anion exchange chromatography Burkholderia thailandensis

Reaction

EC Number Reaction Comment Organism Reaction ID
5.1.3.20 ADP-D-glycero-D-manno-heptose = ADP-L-glycero-D-manno-heptose catalytic one-base reaction mechanism from docking study, overview. The key point in the catalytic process of AGME is the inversion at the C6'' stereocentre of ADP-D,D-Hep mediated by the reversible redox reactions of NADP+/NADPH Burkholderia thailandensis
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5.1.3.20 ADP-D-glycero-D-manno-heptose
-
 Burkholderia thailandensis ADP-L-glycero-D-manno-heptose
-
 r
5.1.3.20 ADP-D-glycero-D-manno-heptose
-
 Burkholderia thailandensis E444 ADP-L-glycero-D-manno-heptose
-
 r

Subunits

EC Number Subunits Comment Organism
5.1.3.20 ? x * 38000, recombinant His6-tagged enzyme, SDS-PAGE Burkholderia thailandensis
5.1.3.20 More enzyme structure comparisons, detailed overview Burkholderia thailandensis

Synonyms

EC Number Synonyms Comment Organism
5.1.3.20 ADP-L-glycero-D-manno-heptose 6-epimerase
-
 Burkholderia thailandensis
5.1.3.20 AGME
-
 Burkholderia thailandensis

Cofactor

EC Number Cofactor Comment Organism Structure
5.1.3.20 NADP+ the enzyme contains a catalytic triad involved in catalyzing hydride transfer with the aid of NADP+ Burkholderia thailandensis
5.1.3.20 NADPH
-
 Burkholderia thailandensis

General Information

EC Number General Information Comment Organism
5.1.3.20 additional information the enzyme contains a catalytic triad involved in catalyzing hydride transfer with the aid of NADP+. The enzyme may recognize their substrate in a lock-and-key fashion, binding analysis, docking study, enzyme structure comparisons, detailed overview Burkholderia thailandensis