Literature summary for 2.7.11.2 extracted from

Knoechel, T.R.; Tucker, A.D.; Robinson, C.M.; Phillips, C.; Taylor, W.; Bungay, P.J.; Kasten, S.A.; Roche, T.E.; Brown, D.G.
Regulatory roles of the N-terminal domain based on crystal structures of human pyruvate dehydrogenase kinase 2 containing physiological and synthetic ligands (2006), Biochemistry, 45, 402-415.

Application

Application	Comment	Organism
medicine	PDHK inhibition provides a route for therapeutic intervention in diabetes and cardiovascular disorders	Homo sapiens

Cloned(Commentary)

Cloned (Comment)	Organism
expression of N-terminally His-tagged residues A8-T399 of isozyme PDHK2 in Trichoplusia ni TN5B1-4 insect cells using the baculovirus infection system	Homo sapiens

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant residues A8-T399 of isozyme PDHK2 free or in complex with ADP and chloroacetate, ATP, Nov3r, Pfz3, or AZ12, hanging drop vapour diffusion method, protein solution contains 10 mg/ml protein, 20 mM Tris-HCl, pH 8.0, 0.15 M NaCl, and 1 mM DTT, versus 0.1 M sodium acetate, pH 5.6-5.8, 6-9% 2-propanol, and 75-125 mM MgCl2, 4°C, 2 weeks, complexing with ligands by soaking of PDHK2 crystals in solutions containing 10 mM ATP, 10 mM ADP and 100 mM dichloroacetate, or 1 mM of Nov3r, Pfz3, or AZ12, cryoprotection by 30-35% glycerol, X-ray diffraction structure determination and analysis at 2.2-3.0 A resolution	Homo sapiens

Crystallization (Comment)

Organism

purified recombinant residues A8-T399 of isozyme PDHK2 free or in complex with ADP and chloroacetate, ATP, Nov3r, Pfz3, or AZ12, hanging drop vapour diffusion method, protein solution contains 10 mg/ml protein, 20 mM Tris-HCl, pH 8.0, 0.15 M NaCl, and 1 mM DTT, versus 0.1 M sodium acetate, pH 5.6-5.8, 6-9% 2-propanol, and 75-125 mM MgCl2, 4°C, 2 weeks, complexing with ligands by soaking of PDHK2 crystals in solutions containing 10 mM ATP, 10 mM ADP and 100 mM dichloroacetate, or 1 mM of Nov3r, Pfz3, or AZ12, cryoprotection by 30-35% glycerol, X-ray diffraction structure determination and analysis at 2.2-3.0 A resolution

Homo sapiens

Inhibitors

Inhibitors	Comment	Organism
ADP	binding site structure, involves Gly319 and Phe318, and K+ ions	Homo sapiens
AZ12	i.e. N-[4-([ethylanilino]sulfonyl)2-methylphenyl]-3,3,3-trifluoro-2-hydroxy-2-methylpropanamide, binding structure, requires K+ for inhibition	Homo sapiens
Dichloroacetate	binds at the pyruvate binding site, binding structure, involves e.g. Arg154	Homo sapiens
Nov3r	i.e. (4-[(2,5)-dimethyl-4-(3,3,3-trifluoro-2-hydroxy-2-methyl-propanoyl)piperazinyl]carbonyl)benzonitrile, binding structure, requires K+ for inhibition	Homo sapiens
Pfz3	i.e. N-(2-aminoethyl)-2-(3-chloro-4-[(4-isopropylbenzyl)oxy]phenyl)acetamide, binding site structure, involves e.g. the R domain, allosteric inhibition mechanism, overview	Homo sapiens

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
mitochondrion	-	Homo sapiens	5739	-

Metals/Ions

Metals/Ions	Comment	Organism	Structure
K+	binds via Gly319, involved in inhibition by ADP, Nov3r, and AZ12, and in binding of ATP	Homo sapiens
Mg2+	-	Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ATP + pyruvate dehydrogenase	Homo sapiens	a small pocket in the N-terminal region of PDHK2 is involved in enzyme regulation, the pocket is formed by residues L53, Y157, Y80, S83, I111, R112, H115, S153, R154, I157, R158, I161	ADP + phosphorylated pyruvate dehydrogenase	-	?
additional information	Homo sapiens	PDK regulates the pyruvate dehydrogenase multienzyme complex activity	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	-	isozyme PDHK2; isozyme PDHK2	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant N-terminally His-tagged residues A8-T399 of isozyme PDHK2 from insect cells by nickel affinity chromatography, the His-tag is cleaved off by thrombin	Homo sapiens

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
ATP + pyruvate dehydrogenase	-	Homo sapiens	ADP + phosphorylated pyruvate dehydrogenase	-	?
ATP + pyruvate dehydrogenase	a small pocket in the N-terminal region of PDHK2 is involved in enzyme regulation, the pocket is formed by residues L53, Y157, Y80, S83, I111, R112, H115, S153, R154, I157, R158, I161	Homo sapiens	ADP + phosphorylated pyruvate dehydrogenase	-	?
additional information	PDK regulates the pyruvate dehydrogenase multienzyme complex activity	Homo sapiens	?	-	?

Subunits

Subunits	Comment	Organism
dimer	PDHK2 is a component of the pyruvate dehydrogenase complex, complex structure	Homo sapiens

Synonyms

Synonyms	Comment	Organism
PDHK2	-	Homo sapiens
pyruvate dehydrogenase kinase 2	-	Homo sapiens

Cofactor

Cofactor	Comment	Organism	Structure
ATP	binding site structure, involves Gly317 and Tyr320, and K+ ions	Homo sapiens