Activating Compound | Comment | Organism | Structure |
---|---|---|---|
Dimethylamine | 9% activation compared to NH4+ | Pseudomonas aeruginosa | |
methylamine | 28% activation compared to NH4+ | Pseudomonas aeruginosa | |
trimethylamine | 5% activation compared to NH4+ | Pseudomonas aeruginosa |
Cloned (Comment) | Organism |
---|---|
gene ppx, recombinant expression of N-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli strains XL10-Gold and BL21-CodonPlus | Pseudomonas aeruginosa |
Protein Variants | Comment | Organism |
---|---|---|
additional information | construction of a truncated enzyme mutants comprising amino acid residues 1-314 of 506 (N-paPpx(1-314)), or residues 1-303 (N-paPpx(1-303)), or residues 315-506 (C-paPpx(315-506)) of paPpx, amplified from Pseudomonas aeruginosa wild-type strain PAO1 chromosomal DNA through PCR. Only paPpx(1-506) and N-paPpx(1-314) are enzymatically active, while C-paPpx(315-506) lacks enzymatic activity | Pseudomonas aeruginosa |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetic analysis of recombinant wild-type and truncated mutant enzymes, polyphosphatase activity (EC 3.6.1.11) and polyphosphate:ADP phosphotransferase activity, overview | Pseudomonas aeruginosa | |
0.00249 | - |
polyphosphate65 | pH 8.0, 37°C, full-length enzyme | Pseudomonas aeruginosa | |
0.00311 | - |
polyphosphate25 | pH 8.0, 37°C, full-length enzyme | Pseudomonas aeruginosa | |
0.00311 | - |
polyphosphate65 | pH 8.0, 37°C, truncated enzyme | Pseudomonas aeruginosa | |
0.00336 | - |
polyphosphate25 | pH 8.0, 37°C, truncated enzyme | Pseudomonas aeruginosa |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Cs+ | activates wild-type, full-length enzyme paPpx(1-506) | Pseudomonas aeruginosa | |
K+ | a nonessential activator of paPpx, presence of K+ does not affect the affinity of the enzyme for Mg2+, activates wild-type, full-length enzyme paPpx(1-506). The activity curve obtained with K+ is sigmoid and reaches its maximum activity at concentrations of 80 mM. Km(app)K+ is 42 mM | Pseudomonas aeruginosa | |
Mg2+ | required, values of Km(app)Mg2+ in paPpx(1-506) and NpaPpx(1-314) are 0.30 mM and 0.28 mM, respectively. The interaction between paPpx(1-506) and Mg2+ occurs in the N-terminal domain | Pseudomonas aeruginosa | |
additional information | behavior of the full-length paPpx(1-506) and N-paPpx(1-314) against different concentration of divalent ions such as Mg2+, Zn2+, Ca2+, and Mn2+ as effectors, in presence of a saturating concentration (0.008 mM) for the substrate polyphosphate65. The activation of both enzyme variants by Mg2+ is similar and shows no inhibition at high concentrations of this ion. The activation by Ca2+ and Mn2+ is negligible. Li+ and Na+ have no effects on enzyme activity, while NH4+, K+, Rb+, and Cs+ are activators of paPpx(1-506). Tetramethylammonium is not an activator of paPpx(1-506) | Pseudomonas aeruginosa | |
NH4+ | activates wild-type, full-length enzyme paPpx(1-506). The activity curve obtained with NH4+ is sigmoid and reaches its maximum activity at concentrations of 30 mM. Km(app)NH4+ is 10 mM | Pseudomonas aeruginosa | |
Rb+ | activates wild-type, full-length enzyme paPpx(1-506) | Pseudomonas aeruginosa | |
Zn2+ | Zn2+ is able to activate the enzyme only 20% compared to Mg2+ | Pseudomonas aeruginosa |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ADP + (phosphate)n | Pseudomonas aeruginosa | - |
ATP + (phosphate)n-1 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas aeruginosa | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21-CodonPlus by nickel affinity chromatography, dialysis, tag cleavage through thrombin, and again dialysis | Pseudomonas aeruginosa |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
15 | - |
recombinant truncated enzyme, pH 8.0, 37°C, ATP formation, in presence of 80 mM K+ | Pseudomonas aeruginosa |
20 | - |
recombinant full-length enzyme, pH 8.0, 37°C, ATP formation, in presence of 80 mM K+ | Pseudomonas aeruginosa |
26.7 | - |
recombinant truncated enzyme, pH 8.0, 37°C, ATP formation, in presence of 25 mM NH4+ | Pseudomonas aeruginosa |
35 | - |
recombinant full-length enzyme, pH 8.0, 37°C, ATP formation, in presence of 25 mM NH4+ | Pseudomonas aeruginosa |
645 | - |
recombinant truncated enzyme, pH 8.0, 37°C, ATP formation, in presence of 5 mM Mg2+ and 80 mM K+ | Pseudomonas aeruginosa |
655 | - |
recombinant truncated enzyme, pH 8.0, 37°C, ATP formation, in presence of 5 mM Mg2+ | Pseudomonas aeruginosa |
715 | - |
recombinant truncated enzyme, pH 8.0, 37°C, ATP formation, in presence of 5 mM Mg2+ and 25 mM NH4+ | Pseudomonas aeruginosa |
3126.7 | - |
recombinant full-length enzyme, pH 8.0, 37°C, ATP formation, in presence of 5 mM Mg2+ and 80 mM K+ | Pseudomonas aeruginosa |
3150 | - |
recombinant full-length enzyme, pH 8.0, 37°C, ATP formation, in presence of 5 mM Mg2+ and 25 mM NH4+ | Pseudomonas aeruginosa |
3223.3 | - |
recombinant full-length enzyme, pH 8.0, 37°C, ATP formation, in presence of 5 mM Mg2+ | Pseudomonas aeruginosa |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ADP + (phosphate)n | - |
Pseudomonas aeruginosa | ATP + (phosphate)n-1 | - |
? | |
ADP + polyphosphate25 | - |
Pseudomonas aeruginosa | ATP + polyphosphate24 | - |
? | |
ADP + polyphosphate65 | - |
Pseudomonas aeruginosa | ATP + polyphosphate64 | - |
? | |
additional information | paPpx is an exopolyphosphatase (EC 3.6.1.11), and is also a polyphosphate:ADP phosphotransferase, and the active site is the same as that one involved in the hydrolase activity | Pseudomonas aeruginosa | ? | - |
- |
Subunits | Comment | Organism |
---|---|---|
? | x * 56419, sequence calculation, wild-type, full-length enzyme | Pseudomonas aeruginosa |
Synonyms | Comment | Organism |
---|---|---|
More | cf. EC 3.6.1.11 | Pseudomonas aeruginosa |
paPpx | - |
Pseudomonas aeruginosa |
polyphosphate: ADP phosphotransferase | - |
Pseudomonas aeruginosa |
polyphosphate:ADP phosphotransferase | - |
Pseudomonas aeruginosa |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Pseudomonas aeruginosa |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.29 | - |
polyphosphate65 | pH 8.0, 37°C, truncated enzyme | Pseudomonas aeruginosa | |
0.31 | - |
polyphosphate25 | pH 8.0, 37°C, truncated enzyme | Pseudomonas aeruginosa | |
3.93 | - |
polyphosphate65 | pH 8.0, 37°C, full-length enzyme | Pseudomonas aeruginosa | |
4.28 | - |
polyphosphate25 | pH 8.0, 37°C, full-length enzyme | Pseudomonas aeruginosa |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Pseudomonas aeruginosa |
General Information | Comment | Organism |
---|---|---|
additional information | homology structural model of full-length paPpx, in closed conformation, and of the N-terminal domain of paPpx in an open state, constructed by comparative modeling, molecular dynamic simulations, overview. Docking study with bound metals and/or ADP defining the N-paPpx(1-314) model in open conformation as receptor, docking with polyphosphate and ADP. Enzyme electrostatic potential calculations. A model of the paPpx N-terminal domain in complex with a polyP chain of 7 residues long and a molecule of ADP explains the phosphotransferase activity through docking techniques, overview | Pseudomonas aeruginosa |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
92.3 | - |
polyphosphate25 | pH 8.0, 37°C, truncated enzyme | Pseudomonas aeruginosa | |
93.3 | - |
polyphosphate65 | pH 8.0, 37°C, truncated enzyme | Pseudomonas aeruginosa | |
1376.2 | - |
polyphosphate25 | pH 8.0, 37°C, full-length enzyme | Pseudomonas aeruginosa | |
1578.3 | - |
polyphosphate65 | pH 8.0, 37°C, full-length enzyme | Pseudomonas aeruginosa |