Cloned (Comment) | Organism |
---|---|
gene coaW, recombinant expression of C-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3) | Staphylococcus aureus |
Crystallization (Comment) | Organism |
---|---|
purified recombinant enzyme SaPanKII in complex with inhibitors N-pentylpantothenamide, N-heptylpantothenamide, N-(5-methoxypentyl)pantothenamide, and N-(benzo[d][1,3]dioxol-5-ylmethyl)pantothenamide, sitting drop vapor diffusion method, 500 nl of 20-30 mg/ml protein in 20 mM Tris-HCl, pH 8.0, 200 mM NaCl, and 10 mM dithiothreitol, and addition of 1.1% w/v cyclohexylethanoyl-N-hydroxyethylglucamide, ligand, and ATP, is mixed with 500 nl of well solution containing 25-35% PEG 4000, 0.2 M MgCl2, and 0.1 M Tris, pH 8.5, X-ray diffraction structure determination and analysis at 1.40-1.80 A resolution | Staphylococcus aureus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | the potent antistaphylococcal activity of N-substituted pantothenamides (PanAms) exhibit inhibition of Staphylococcus aureus atypical type II pantothenate kinase (SaPanKII). The PanAms are phosphorylated by SaPanKII but remain bound at the active site. This occurs primarily through interactions with Tyr240' and Thr172'. Kinetic analysis shows a strong correlation between kcat (slow PanAm turnover) and IC50 (inhibition of pantothenate phosphorylation) values, suggesting that SaPanKII inhibition occurs via a delay in product release. The two PanAms, N-(benzo[d][1,3]dioxol-5-ylmethyl)pantothenamide and N-(5-methoxypentyl)pantothenamide, effectively combine both hydrogen bonding and hydrophobic interactions, resulting in the most potent SaPanKII inhibition described to date. Design and synthesis of a series of N-substituted pantothenamides, analysis of the PanAm binding mode and interaction with the N-substituent binding site, structure-activity realtionships, MIC values, overview | Staphylococcus aureus | |
N-(2-(benzo[d][1,3]dioxol-5-yl)ethyl)pantothenamide | - |
Staphylococcus aureus | |
N-(3-methoxyphenethyl)pantothenamide | - |
Staphylococcus aureus | |
N-(5-methoxypentyl)pantothenamide | - |
Staphylococcus aureus | |
N-(benzo[d][1,3]dioxol-5-ylmethyl)pantothenamide | - |
Staphylococcus aureus | |
N-heptylpantothenamide | - |
Staphylococcus aureus | |
N-pentylpantothenamide | - |
Staphylococcus aureus | |
N-phenethylpantothenamide | - |
Staphylococcus aureus | |
pantothenamide | - |
Staphylococcus aureus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Staphylococcus aureus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + (R)-pantothenate | Staphylococcus aureus | - |
ADP + (R)-4'-phosphopantothenate | - |
? | |
ATP + (R)-pantothenate | Staphylococcus aureus RN4220 | - |
ADP + (R)-4'-phosphopantothenate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Staphylococcus aureus | A0A167Z3Z6 | - |
- |
Staphylococcus aureus RN4220 | A0A167Z3Z6 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant C-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, desalting gel filtration, anion exchange chromatography, gel filtration, and ultrafiltration | Staphylococcus aureus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + (R)-pantothenate | - |
Staphylococcus aureus | ADP + (R)-4'-phosphopantothenate | - |
? | |
ATP + (R)-pantothenate | - |
Staphylococcus aureus RN4220 | ADP + (R)-4'-phosphopantothenate | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | the subunit of the homodimers is composed of two domains (domains I and II) and the binding cap (residues 153-173), with a dimerization interface along helix alpha6 | Staphylococcus aureus |
Synonyms | Comment | Organism |
---|---|---|
coaW | - |
Staphylococcus aureus |
PanK | - |
Staphylococcus aureus |
type III pantothenate kinase | - |
Staphylococcus aureus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Staphylococcus aureus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.6 | - |
assay at | Staphylococcus aureus |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.00013 | - |
pH 7.6, 25°C | Staphylococcus aureus | N-(5-methoxypentyl)pantothenamide | |
0.00046 | - |
pH 7.6, 25°C | Staphylococcus aureus | N-(benzo[d][1,3]dioxol-5-ylmethyl)pantothenamide | |
0.006 | - |
pH 7.6, 25°C | Staphylococcus aureus | N-pentylpantothenamide | |
0.01 | - |
pH 7.6, 25°C | Staphylococcus aureus | N-(3-methoxyphenethyl)pantothenamide | |
0.018 | - |
pH 7.6, 25°C | Staphylococcus aureus | N-heptylpantothenamide | |
0.02 | - |
pH 7.6, 25°C | Staphylococcus aureus | N-(2-(benzo[d][1,3]dioxol-5-yl)ethyl)pantothenamide | |
0.036 | - |
pH 7.6, 25°C | Staphylococcus aureus | N-phenethylpantothenamide |
General Information | Comment | Organism |
---|---|---|
malfunction | the potent antistaphylococcal activity of N-substituted pantothenamides (PanAms) exhibit inhibition of Staphylococcus aureus's atypical type II pantothenate kinase (SaPanKII), the first enzyme of coenzyme A biosynthesis. The mechanism of action follows from SaPanKII having a binding mode for PanAms that is distinct from those of other PanKs. Molecular interactions responsible for PanAm inhibitory activity, overview. The PanAms are phosphorylated by SaPanKII but remain bound at the active site, SaPanKII inhibition occurs via a delay in product release | Staphylococcus aureus |
metabolism | CoA biosynthesis and salvage pathways, overview. The Staphylococcus aureus atypical type II pantothenate kinase (SaPanKII) is active in bothe pathways | Staphylococcus aureus |