Literature summary for 2.7.1.33 extracted from

Ivey, R.A.; Zhang, Y.M.; Virga, K.G.; Hevener, K.; Lee, R.E.; Rock, C.O.; Jackowski, S.; Park, H.W.
The structure of the pantothenate kinase-ADP-pantothenate ternary complex reveals the relationship between the binding sites for substrate, allosteric regulator, and antimetabolites (2004), J. Biol. Chem., 279, 35622-35629.

Cloned(Commentary)

Cloned (Comment)	Organism
gene coaA, expression in strain BL21(DE3)	Escherichia coli

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant enzyme in a ternary complex with ADP and pantothenate, hanging drop vapour diffusion method, 30 mg/ml protein in 20 mM Tris-HCl, pH 8.0, 1 mM DTT, 1 mM EDTA, incubated overnight with 30 mM ADP, 30 mM pantothenate, and 30 mM magnesium nitrate, mixing in equal volumes with reservoir solution containing 11% PEG 3350, 0.2 M sodium citrate, pH 8.2, at 18°C, 1 week, X-ray diffraction structure determination and analysis at 2.2 A resolution, molecular modeling of N-alkylpantothenamides, growth-inhibitory anti-metabolites	Escherichia coli

Crystallization (Comment)

Organism

purified recombinant enzyme in a ternary complex with ADP and pantothenate, hanging drop vapour diffusion method, 30 mg/ml protein in 20 mM Tris-HCl, pH 8.0, 1 mM DTT, 1 mM EDTA, incubated overnight with 30 mM ADP, 30 mM pantothenate, and 30 mM magnesium nitrate, mixing in equal volumes with reservoir solution containing 11% PEG 3350, 0.2 M sodium citrate, pH 8.2, at 18°C, 1 week, X-ray diffraction structure determination and analysis at 2.2 A resolution, molecular modeling of N-alkylpantothenamides, growth-inhibitory anti-metabolites

Escherichia coli

Inhibitors

Inhibitors	Comment	Organism
coenzyme A	allosteric regulator, feedback inhibition	Escherichia coli
additional information	no inhibition by hopantenate	Escherichia coli
N-heptylpantothenamide	competitive to pantothenate	Escherichia coli
N-pentylpantothenamide	competitive to pantothenate	Escherichia coli

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.041	-	(R)-pantothenate	pH 7.5, 37°C, recombinant His-tagged enzyme	Escherichia coli
0.124	-	N-heptylpantothenamide	pH 7.5, 37°C, recombinant His-tagged enzyme	Escherichia coli
0.14	-	N-pentylpantothenamide	pH 7.5, 37°C, recombinant His-tagged enzyme	Escherichia coli

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	coordinated by the nucleotide beta- and gamma-phosphates and the side chains of Ser102 and Glu199, required for ATP but not for ADP binding	Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ATP + (R)-pantothenate	Escherichia coli	first step in coenzyme A biosynthesis, the enzyme has a regulatory function in the pathway	ADP + (R)-4'-phosphopantothenate	-	?

Organism

Organism	UniProt	Comment	Textmining
Escherichia coli	P0A6I3	gene coaA	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant enzyme from strain BL21(DE3) by ion exchange and hydrophobic interaction chromatography, and gel filtration, recombinant His-tagged enzyme from strain BL21(DE3) by nickel affinity chromatography	Escherichia coli

Reaction

Reaction	Comment	Organism	Reaction ID
ATP + (R)-pantothenate = ADP + (R)-4'-phosphopantothenate	active site structure, mechanism, pantothenate binding site structure, the reaction proceeds by a concerted mechanism that involves a dissociative transition state, although the negative charge neutralization of the gamma-phosphate by Arg243, Lys101, and Mg2+ coupled with hydrogen bonding of the C1 of pantothenate to Asp127 suggests different interpretations of the phosphoryl transfer mechanism of pantothenate kinase	Escherichia coli	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
ATP + (R)-pantothenate	first step in coenzyme A biosynthesis, the enzyme has a regulatory function in the pathway	Escherichia coli	ADP + (R)-4'-phosphopantothenate	-	?
ATP + (R)-pantothenate	pantothenate binding site structure involving residues E249, Y262, F247, F259, Y258, and F244, located at the distal end of a large surface groove, induced fit binding mechanism, overview	Escherichia coli	ADP + (R)-4'-phosphopantothenate	-	?
ATP + N-alkylpantothenamides	growth-inhibiting anti-metabolite, modeling into the active site structure	Escherichia coli	ADP + ?	-	?
ATP + N-heptylpantothenamide	-	Escherichia coli	ADP + N-heptylpantothenamide 4-phosphate	-	?
ATP + N-pentylpantothenamide	-	Escherichia coli	ADP + N-pentylpantothenamide 4-phosphate	-	?
additional information	no acivity with hopantenate, formation of a pantothenate kinase-ADP-pantothenate ternary complex	Escherichia coli	?	-	?

Subunits

Subunits	Comment	Organism
More	formation of a pantothenate kinase-ADP-pantothenate ternary complex, structure determination, pantothenate binding to the enzyme induces a significant conformational change in amino acids 243263, which form a lid that folds over the open pantothenate binding groove	Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Escherichia coli

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	assay at	Escherichia coli

Cofactor

Cofactor	Comment	Organism	Structure
ATP	three-dimensional binding site structure, Arg243 is involved	Escherichia coli