Cloned (Comment) | Organism |
---|---|
gene adk, recombinant expression of GST-tagged enzyme in Escherichia coli strain BL21(DE3) | Streptococcus pneumoniae |
Crystallization (Comment) | Organism |
---|---|
purified recombinant enzyme in ligand-free and inhibitor Ap5A-bound states, hanging drop vapour diffusion method, mixing of 0.0025 ml of 10 mg/ml protein in 50 mM Tris-HCl, pH 7.5, 50-150 mM NaCl, 5 mM MgCl2, with 0.0025 ml reservoir solution containing 1.0 M sodium citrate, 0.1 M HEPES, pH 7.5 for the ligand-free crystals and 0.2 M sodium acetate, pH 7.2, 30% PEG 8000 for the inhibitor-bound crystals, 22°C, X-ray diffraction structure determination and analysis at 1.96 and 1.65 A resolution, respectively. SpAdK can crystallize promiscuously in different forms, and the open structure is flexible in conformation | Streptococcus pneumoniae |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
P1,P5-bis(adenosine-5'-)pentaphosphate | i.e. Ap5A, a two substrate-mimicking inhibitor, binding affinity of SpAdK to Ap5A is determined by isothermal titration calorimetry | Streptococcus pneumoniae |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Streptococcus pneumoniae |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
24000 | - |
- |
Streptococcus pneumoniae |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + AMP | Streptococcus pneumoniae | - |
2 ADP | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptococcus pneumoniae | Q04ML5 | gene adk | - |
Purification (Comment) | Organism |
---|---|
recombinant GST-tagged enzyme from Escherichia coli strain BL21(DE3) by glutathione affinity chromatography and gel filtration, the GST moiety is cleaved using tobacco etch virus protease | Streptococcus pneumoniae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + AMP | - |
Streptococcus pneumoniae | 2 ADP | - |
r |
Subunits | Comment | Organism |
---|---|---|
? | x * 24000, detagged recombinant enzyme, SDS-PAGE, x * 23721, sequence calculation | Streptococcus pneumoniae |
Synonyms | Comment | Organism |
---|---|---|
ADK | - |
Streptococcus pneumoniae |
General Information | Comment | Organism |
---|---|---|
additional information | the ligand-free enzyme structure reveals an open conformation | Streptococcus pneumoniae |
physiological function | adenylate kinases play a critical role in intercellular homeostasis by the interconversion of ATP and AMP to two ADP molecules | Streptococcus pneumoniae |