Cloned (Comment) | Organism |
---|---|
gene adk, recombinant expression of wild-type adk gene in fucose-inducible strains rescues a growth defect, but expression of the Arg89 mutant does not. Recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) | Streptococcus pneumoniae |
Crystallization (Comment) | Organism |
---|---|
purified detagged recombinant enzyme free or in complex with inhibitor Ap5A, hanging drop vapour diffusion method, mixing of 0.0025 ml of 18 mg/ml protein in 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, and 1 mM MgCl2, with 0.0025 ml of reservoir solution containing 2.0 M (NH4)2SO4, 0.1 M CHES, pH 9.5, 0.2 M Li2SO4, and 0.1 M CsCl2 for the ligand-free enzyme, and 0.1 M sodium acetate, 0.1 M sodium acetate, pH 4.6, 30% PEG 8000, and 50 mM NaF for the inhibitor-bound enzyme, 22°C, X-ray diffraction structure determination and analysis at 1.7 and 1.48 A resolution, respectively, molecular replacement using structures Marinibacillus marinus PDB ID 3FB4 and Burkholderia pseudomallei PDB ID 3GMT as search models | Streptococcus pneumoniae |
Protein Variants | Comment | Organism |
---|---|---|
additional information | construction of a conditional mutant using fucose inducible adk promoter, the D39 fucose-inducible adk strain | Streptococcus pneumoniae |
R89A | site-directed mutagenesis, inactive active site mutant | Streptococcus pneumoniae |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
P1,P5-bis(adenosine-5'-)pentaphosphate | i.e. Ap5A | Streptococcus pneumoniae |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | 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 | serotype 2, gene adk | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography. The tag is cleaved using tobacco etch virus protease, followed by dialysis, gel filtration, and ultrafiltration | Streptococcus pneumoniae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + AMP | - |
Streptococcus pneumoniae | 2 ADP | - |
r |
Synonyms | Comment | Organism |
---|---|---|
ADK | - |
Streptococcus pneumoniae |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Streptococcus pneumoniae |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.2 | - |
assay at | Streptococcus pneumoniae |
General Information | Comment | Organism |
---|---|---|
malfunction | rrecombinant expression of wild-type adk gene in fucose-inducible strains rescues a growth defect, but expression of the Arg89 mutant does not. Lack of functional SpAdK causes a growth defect in vivo, SpAdK deficiency abolishes pneumococcal growth | Streptococcus pneumoniae |
additional information | Arg89 is a key active site residue, enzyme structure comparisons | Streptococcus pneumoniae |
physiological function | SpAdK plays an essential role in pneumococcal growth and ATP synthesis, the enzyme is essential for growth through its catalytic activity, it controls cell growth via cellular energy homeostasis. SpAdK increases total cellular ATP levels regulated by fucose in the fucose-inducible strain | Streptococcus pneumoniae |