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Literature summary extracted from

  • Sgraja, T.; Alphey, M.S.; Ghilagaber, S.; Marquez, R.; Robertson, M.N.; Hemmings, J.L.; Lauw, S.; Rohdich, F.;Bacher, A.; Eisenreich, W.; Illarionova, V.; Hunter, W.N.
    Characterization of Aquifex aeolicus 4-diphosphocytidyl-2C-methyl-d-erythritol kinase - ligand recognition in a template for antimicrobial drug discovery (2008), FEBS J., 275, 2779-2794.
    View publication on PubMedView publication on EuropePMC

Application

EC Number Application Comment Organism
2.7.1.148 drug development potential target for anti-infective drugs Aquifex aeolicus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.1.148 The putative ispE gene from Aquifex aeolicus is placed under the control of the T7 promoter and lac operator in the hyperexpression plasmid pET-15b, and heat transformed into Escherichia coli BL21 (DE3). Aquifex aeolicus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.7.1.148 AaIspE is crystallized by vapor diffusion. Ordered and reproducible crystals of AaIspE are obtained and multiple-wavelength anomalous dispersion methods are applied to obtain the initial phases. 3 medium-resolution complex crystal structures are determined. The crystals are isomorphous with two molecules in the asymmetric unit. Aquifex aeolicus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.1.148 0.121
-
 4-diphosphocytidyl-2C-methyl-D-erythritol
-
 Aquifex aeolicus
2.7.1.148 0.222
-
 ATP
-
 Aquifex aeolicus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.1.148 Ca2+ activity rate 8% compared to Mg2+ Aquifex aeolicus
2.7.1.148 Co2+ activity rate 52% compared to Mg2+ Aquifex aeolicus
2.7.1.148 Cu2+ activity rate 40% compared to Mg2+ Aquifex aeolicus
2.7.1.148 Fe2+ activity rate 16% compared to Mg2+ Aquifex aeolicus
2.7.1.148 Mg2+ activity rate 100% Aquifex aeolicus
2.7.1.148 Mn2+ activity rate 94% compared to Mg2+ Aquifex aeolicus
2.7.1.148 Ni2+ activity rate 11% compared to Mg2+ Aquifex aeolicus
2.7.1.148 Zn2+ activity rate 14% compared to Mg2+ Aquifex aeolicus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.1.148 30000
-
 by gel-filtration chromatography Aquifex aeolicus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.1.148 4-diphosphocytidyl-2C-methyl-D-erythritol + ATP Aquifex aeolicus non-mevalonate pathway of isoprenoid precursor synthesis 4-diphosphocytidyl-2C-methyl-D-erythritol 2-phosphate + ADP
-
 ir

Organism

EC Number Organism UniProt Comment Textmining
2.7.1.148 Aquifex aeolicus O67060
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.1.148 The gene is preceded by a His6 tag to enable purification of the recombinant protein via metal-chelating affinity chromatography. The polyhistidine tag is removed by thrombin-mediated proteolysis, followed by purification with anion-exchange chromatography. Purity of the sample is assessed by SDS-PAGE and MALDI-TOF MS. Aquifex aeolicus

Source Tissue

EC Number Source Tissue Comment Organism Textmining
2.7.1.148 cell culture cloned in Escherichia coli Aquifex aeolicus
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.7.1.148 additional information
-
 Activation energy is 64.2 kJ/mol Aquifex aeolicus
2.7.1.148 1.5
-
 in presence of 2 mM Mg2+ or Mn2+ Aquifex aeolicus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.1.148 4-diphosphocytidyl-2C-methyl-D-erythritol + ATP non-mevalonate pathway of isoprenoid precursor synthesis Aquifex aeolicus 4-diphosphocytidyl-2C-methyl-D-erythritol 2-phosphate + ADP
-
 ir

Subunits

EC Number Subunits Comment Organism
2.7.1.148 monomer Gel-filtration and analytical ultracentrifugation indicate that AaIspE is a monomer in solution Aquifex aeolicus

Synonyms

EC Number Synonyms Comment Organism
2.7.1.148 4-diphosphocytidyl-2C-methyl-D-erythritol kinase
-
 Aquifex aeolicus
2.7.1.148 AaIspE
-
 Aquifex aeolicus
2.7.1.148 CMK
-
 Aquifex aeolicus
2.7.1.148 IspE
-
 Aquifex aeolicus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.1.148 60
-
-
 Aquifex aeolicus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.1.148 8.5
-
-
 Aquifex aeolicus

Cofactor

EC Number Cofactor Comment Organism Structure
2.7.1.148 ATP
-
 Aquifex aeolicus