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Literature summary extracted from

  • Thach, T.T.; Luong, T.T.; Lee, S.; Rhee, D.K.
    Adenylate kinase from Streptococcus pneumoniae is essential for growth through its catalytic activity (2014), FEBS open bio, 4, 672-682.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.4.3 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 (Commentary)

EC Number Crystallization (Comment) Organism
2.7.4.3 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

EC Number Protein Variants Comment Organism
2.7.4.3 additional information construction of a conditional mutant using fucose inducible adk promoter, the D39 fucose-inducible adk strain Streptococcus pneumoniae
2.7.4.3 R89A site-directed mutagenesis, inactive active site mutant Streptococcus pneumoniae

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.7.4.3 P1,P5-bis(adenosine-5'-)pentaphosphate i.e. Ap5A Streptococcus pneumoniae

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.4.3 Mg2+ required Streptococcus pneumoniae

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.4.3 ATP + AMP Streptococcus pneumoniae
-
 2 ADP
-
 r

Organism

EC Number Organism UniProt Comment Textmining
2.7.4.3 Streptococcus pneumoniae Q04ML5 serotype 2, gene adk
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.4.3 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 and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.4.3 ATP + AMP
-
 Streptococcus pneumoniae 2 ADP
-
 r

Synonyms

EC Number Synonyms Comment Organism
2.7.4.3 ADK
-
 Streptococcus pneumoniae

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.4.3 37
-
 assay at Streptococcus pneumoniae

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.4.3 7.2
-
 assay at Streptococcus pneumoniae

General Information

EC Number General Information Comment Organism
2.7.4.3 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
2.7.4.3 additional information Arg89 is a key active site residue, enzyme structure comparisons Streptococcus pneumoniae
2.7.4.3 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