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Literature summary extracted from

  • Yoshida, A.; Tomita, T.; Kurihara, T.; Fushinobu, S.; Kuzuyama, T.; Nishiyama, M.
    Structural Insight into concerted inhibition of alpha 2 beta 2-type aspartate kinase from Corynebacterium glutamicum (2007), J. Mol. Biol., 368, 521-536.
    View publication on PubMed

Application

EC Number Application Comment Organism
2.7.2.4 additional information AK has biological importance, as a target candidate for developing new antifungal and antibacterial compounds, because mammals cannot biosynthesize lysine. Corynebacterium glutamicum

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.2.4 expression in Escherichia coli BL21-CodonPlus(DE3)-RIL cells with pET-26b(+) vector Corynebacterium glutamicum
2.7.2.4 into vector pET-26b(+), introduced into Escherichia coli BL21-CodonPlus(DE3)-RIL cells Corynebacterium glutamicum

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.7.2.4 by the hanging-drop vapor-diffusion method, crystal structure of the regulatory subunit of AK at 1.58 A resolution in the Thr-binding form, regulatory subunit contains two ACT domain motifs per monomer and is arranged as a dimer, regulatory subunit is a monomer in the absence of Thr but becomes a dimer by adding Thr Corynebacterium glutamicum
2.7.2.4 the structure of the regulatory subiunit AKbeta complexed with threonine is determined at 1.58 A resolution, 0.2 M ammonium sulfate, 0.1 M citrate, 36%(w/v) PEG 4000, 10 mM threonine, pH 5.0, vapor diffusion, hanging drop, temperature 293 K, space group C 2 (C 1 2 1) Corynebacterium glutamicum

Protein Variants

EC Number Protein Variants Comment Organism
2.7.2.4 E114A changes in the inhibitory profile upon addition of Thr, monomer even with the addition of Thr Corynebacterium glutamicum
2.7.2.4 E114A changes in the inhibitory profile upon addition of threonine, monomer even with the addition of Thr Corynebacterium glutamicum
2.7.2.4 F115A changes in the inhibitory profile upon addition of Thr, pentamer or hexamer in the presence and in the absence of Thr Corynebacterium glutamicum
2.7.2.4 F115A changes in the inhibitory profile upon addition of threonine, pentamer or hexamer in the presence and in the absence of Thr Corynebacterium glutamicum
2.7.2.4 G110A mutant enzyme shows normal and negligible response to Thr and Lys, dimerized by Thr Corynebacterium glutamicum
2.7.2.4 G28A changes in the inhibitory profile upon addition of Thr Corynebacterium glutamicum
2.7.2.4 G28A changes in the inhibitory profile upon addition of threonine Corynebacterium glutamicum
2.7.2.4 K106A mutant showing change in Lys response Corynebacterium glutamicum
2.7.2.4 M105A mutant showing change in Lys response Corynebacterium glutamicum
2.7.2.4 N50A mutant showing change in Lys response Corynebacterium glutamicum
2.7.2.4 Q49A changes in the inhibitory profile upon addition of Thr, monomer even with the addition of Thr Corynebacterium glutamicum
2.7.2.4 Q49A changes in the inhibitory profile upon addition of threonine, monomer even with the addition of Thr Corynebacterium glutamicum
2.7.2.4 T112A changes in the inhibitory profile upon addition of Thr, monomer even with the addition of Thr Corynebacterium glutamicum
2.7.2.4 T112A changes in the inhibitory profile upon addition of threonine, monomer even with the addition of Thr Corynebacterium glutamicum
2.7.2.4 V111A changes in the inhibitory profile upon addition of Thr, pentamer or hexamer in the presence and in the absence of Thr Corynebacterium glutamicum
2.7.2.4 V111A changes in the inhibitory profile upon addition of threonine, pentamer or hexamer in the presence and in the absence of Thr Corynebacterium glutamicum
2.7.2.4 V51A mutant showing change in Lys response Corynebacterium glutamicum

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.7.2.4 L-lysine AK is inhibited moderately by Lys, the enzyme is inhibited dramatically by simultaneous addition of Lys and Thr, dimerization of the regulatory subunit induced by Thr binding is a key step in the inhibitory mechanism of AK; feedback inhibition, inhibits moderately, simultaneous addition of lysine and threonine inhibits dramatically Corynebacterium glutamicum
2.7.2.4 L-threonine Thr alone has no effect on the inhibitory profile, the enzyme is inhibited dramatically by simultaneous addition of Lys and Thr; threonine alone has no effect, simultaneous addition of lysine and threonine inhibits dramatically Corynebacterium glutamicum
2.7.2.4 additional information dimerization of the regulatory subunit by Thr binding is the critical step of inhibition Corynebacterium glutamicum

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.2.4 Mg2+
-
 Corynebacterium glutamicum

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.2.4 20200
-
 AKbeta analytical ultracentrifugation, absence of threonine Corynebacterium glutamicum
2.7.2.4 23000
-
 AKbeta, gel filtration Corynebacterium glutamicum
2.7.2.4 23000
-
 AKbeta gel-filtration chromatography, absence of additives, somewhat larger than the mass of a monomer, addition of lysine causes no change Corynebacterium glutamicum
2.7.2.4 33300
-
 AKbeta gel-filtration chromatography, addition of threonine induces dimerization Corynebacterium glutamicum
2.7.2.4 33800
-
 AKbeta in presence with 5 mM L-threonine, gel filtration Corynebacterium glutamicum
2.7.2.4 36000
-
 AKbeta analytical ultracentrifugation, presence of threonine Corynebacterium glutamicum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.2.4 ATP + L-aspartate Corynebacterium glutamicum
-
 ADP + 4-phospho-L-aspartate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.7.2.4 Corynebacterium glutamicum
-
-
-
2.7.2.4 Corynebacterium glutamicum P26512 expression in Escherichia coli BL21-CodonPlus(DE3)-RIL cells with pET-26b(+) vector
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.2.4 by Ni2+-affinity chromatography and gel filtration Corynebacterium glutamicum
2.7.2.4 Ni2+-affinity with Ni2+-NTA resin and gel-filtration chromatogrphy Corynebacterium glutamicum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.2.4 ATP + L-aspartate
-
 Corynebacterium glutamicum ADP + 4-phospho-L-aspartate
-
 ?

Subunits

EC Number Subunits Comment Organism
2.7.2.4 homodimer regulatory subunit of an alpha2beta2-type AK, crystallography Corynebacterium glutamicum
2.7.2.4 homodimer AKbeta is the regulatory subunit of the alpha2beta2 heterotetramer and contains two ACT domain motifs per monomer Corynebacterium glutamicum
2.7.2.4 More The regulatory subunit of AK is a monomer in the absence of Thr but becomes a dimer by adding Thr Corynebacterium glutamicum

Synonyms

EC Number Synonyms Comment Organism
2.7.2.4 AK
-
 Corynebacterium glutamicum
2.7.2.4 aspartate kinase
-
 Corynebacterium glutamicum