Cloned (Comment) | Organism |
---|---|
gene coaA, expression in strain BL21(DE3) | 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 | Comment | Organism | Structure |
---|---|---|---|
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 Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
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 | 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 | 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 | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P0A6I3 | gene coaA | - |
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 | 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 |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
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 | 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 Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | three-dimensional binding site structure, Arg243 is involved | Escherichia coli |