Literature summary for 2.7.2.4 extracted from

Liu, X.; Pavlovsky, A.G.; Viola, R.E.
The structural basis for allosteric inhibition of a threonine-sensitive aspartokinase (2008), J. Biol. Chem., 283, 16216-16225.

Application

Application	Comment	Organism
additional information	The absence of the aspartate biosynthetic pathway in humans makes it a good target for new pesticides and antibiotics.	Methanocaldococcus jannaschii

Cloned(Commentary)

Cloned (Comment)	Organism
pET-41a vector and Rosetta (DE3) Escherichia coli cells for protein expression	Methanocaldococcus jannaschii

Crystallization (Commentary)

Crystallization (Comment)	Organism
3C20, 14% PEG4000, 0.1 M Tris, 0.8 M ammonium formate, pH 8.0, vapor diffusion, hanging drop, temperature 293 K, space group C2221, 3C1N, 0.2 M ammonium iodide, 2.2 M ammonium sulfate, pH 5.0, vapor diffusion, hanging drop, temperature 293 K, space group P212121, 3C1M, 14% PEG4000, 100 mM Tris, 800 mM ammonium formate, pH 8.0, vapor diffusion, hanging drop, temperature 293 K, space group P212121, the structure is determined under three different transition states: ternary complex with MgAMP-PNP and L-aspartate, binary complex with L-aspartate, and binary complex in the presence of its allosteric inhibitor L-threonine	Methanocaldococcus jannaschii

Crystallization (Comment)

Organism

3C20, 14% PEG4000, 0.1 M Tris, 0.8 M ammonium formate, pH 8.0, vapor diffusion, hanging drop, temperature 293 K, space group C2221, 3C1N, 0.2 M ammonium iodide, 2.2 M ammonium sulfate, pH 5.0, vapor diffusion, hanging drop, temperature 293 K, space group P212121, 3C1M, 14% PEG4000, 100 mM Tris, 800 mM ammonium formate, pH 8.0, vapor diffusion, hanging drop, temperature 293 K, space group P212121, the structure is determined under three different transition states: ternary complex with MgAMP-PNP and L-aspartate, binary complex with L-aspartate, and binary complex in the presence of its allosteric inhibitor L-threonine

Methanocaldococcus jannaschii

Inhibitors

Inhibitors	Comment	Organism	Structure
L-threonine	mechanism for allosteric regulation in which the domain movements induced by L-threonine binding causes displacement of the substrates from the enzyme, resulting in a relaxed, inactive conformation	Methanocaldococcus jannaschii

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	-	Methanocaldococcus jannaschii

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ATP + L-aspartate	Methanocaldococcus jannaschii	L-aspartate binds to this recombinant enzyme in two different orientations	ADP + 4-phospho-L-aspartate	-	?

Organism

Organism	UniProt	Comment	Textmining
Methanocaldococcus jannaschii	Q57991	Methanococcus jannaschii has only a single, monofunctional form of AK; pET-41a vector and Rosetta (DE3) Escherichia coli cells for protein expression	-

Purification (Commentary)

Purification (Comment)	Organism
two successive chromatography steps: first a high resolution anionic exchange resin, followed by gel filtration column, the resulting protein is shown by SDS-PAGE to be more than 99% pure, concentration to 30 mg/ml by using a 10000 molecular weight cut-off Amicon concentrator	Methanocaldococcus jannaschii

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
ATP + L-aspartate	L-aspartate binds to this recombinant enzyme in two different orientations	Methanocaldococcus jannaschii	ADP + 4-phospho-L-aspartate	-	?

Subunits

Subunits	Comment	Organism
tetramer	dimer of dimers	Methanocaldococcus jannaschii

Synonyms

Synonyms	Comment	Organism
aspartokinase	-	Methanocaldococcus jannaschii