Literature summary for 2.7.2.4 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.

Search for more literature for 2.7.2.4

Application

Application	Comment	Organism
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)

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

Crystallization (Commentary)

Crystallization (Comment)	Organism
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
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

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

Inhibitors

Inhibitors	Comment	Organism	Structure
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
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
additional information	dimerization of the regulatory subunit by Thr binding is the critical step of inhibition	Corynebacterium glutamicum

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	-	Corynebacterium glutamicum

Molecular Weight [Da]

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

Natural Substrates/ Products (Substrates)

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

Organism

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

Purification (Commentary)

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

Substrates and Products (Substrate)

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

Subunits

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

Synonyms

Synonyms	Comment	Organism
AK	-	Corynebacterium glutamicum
aspartate kinase	-	Corynebacterium glutamicum

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Release 2023.1 (January 2023)