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Literature summary for 2.7.1.148 extracted from
- 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.
Application
| Application | Comment | Organism |
|---|---|---|
| drug development | potential target for anti-infective drugs | Aquifex aeolicus |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| 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)
| Crystallization (Comment) | Organism |
|---|---|
| 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]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.121 | - |
4-diphosphocytidyl-2C-methyl-D-erythritol | - |
Aquifex aeolicus |  |
| 0.222 | - |
ATP | - |
Aquifex aeolicus | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Ca2+ | activity rate 8% compared to Mg2+ | Aquifex aeolicus | |
| Co2+ | activity rate 52% compared to Mg2+ | Aquifex aeolicus | |
| Cu2+ | activity rate 40% compared to Mg2+ | Aquifex aeolicus | |
| Fe2+ | activity rate 16% compared to Mg2+ | Aquifex aeolicus | |
| Mg2+ | activity rate 100% | Aquifex aeolicus | |
| Mn2+ | activity rate 94% compared to Mg2+ | Aquifex aeolicus | |
| Ni2+ | activity rate 11% compared to Mg2+ | Aquifex aeolicus | |
| Zn2+ | activity rate 14% compared to Mg2+ | Aquifex aeolicus | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 30000 | - |
by gel-filtration chromatography | Aquifex aeolicus |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 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
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Aquifex aeolicus | O67060 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| 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
| Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|
| cell culture | cloned in Escherichia coli | Aquifex aeolicus | - |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| additional information | - |
Activation energy is 64.2 kJ/mol | Aquifex aeolicus |
| 1.5 | - |
in presence of 2 mM Mg2+ or Mn2+ | Aquifex aeolicus |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 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
| Subunits | Comment | Organism |
|---|---|---|
| monomer | Gel-filtration and analytical ultracentrifugation indicate that AaIspE is a monomer in solution | Aquifex aeolicus |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| 4-diphosphocytidyl-2C-methyl-D-erythritol kinase | - |
Aquifex aeolicus |
| AaIspE | - |
Aquifex aeolicus |
| CMK | - |
Aquifex aeolicus |
| IspE | - |
Aquifex aeolicus |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 60 | - |
- |
Aquifex aeolicus |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8.5 | - |
- |
Aquifex aeolicus |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| ATP | - |
Aquifex aeolicus | |
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