Cloned (Comment) | Organism |
---|---|
two recombinant forms of the Porphyromonas gingivalis PAD, a truncated form with a 43-amino acid N-terminal deletion and the full-length form of PAD, are expressed in Escherichia coli BL21 Star (DE3)pLysS cells | Porphyromonas gingivalis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Cd2+ | 53% inhibition at 1 mM Cd2+ | Porphyromonas gingivalis | |
Zn2+ | at 1 mM Zn2+, the enzyme activity is inhibited by 26% | Porphyromonas gingivalis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.01 | 0.02 | N-alpha-benzoyl-L-arginine | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis | |
0.021 | - |
bradykinin-L-Arg | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis | |
0.033 | - |
bradykinin antagonist-L-Arg | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis | |
0.07 | - |
L-Pro-L-Arg-L-Phe | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis | |
0.089 | - |
L-Pro-L-Phe-L-Arg | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis | |
1.2 | - |
L-Arg | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis | |
1.5 | - |
Nomega-hydroxy-L-arginine | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis | |
1.9 | - |
D-Arg | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis | |
4.7 | - |
homoarginine | - |
Porphyromonas gingivalis | |
6 | - |
L-canavanine-L-Arg | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis | |
6.9 | - |
L-Arginine hydroxamate | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis | |
18 | - |
benzoyl-L-arginine ethyl ester | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis | |
31 | - |
agmatine-L-Arg | - |
Porphyromonas gingivalis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
additional information | PAD is not a metalloenzyme | Porphyromonas gingivalis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
51000 | - |
purified recombinant truncated PAD (missing 43 N-terminal amino acid residues but including Xpress and poly-His tags), SDS-PAGE | Porphyromonas gingivalis |
53000 | - |
active enzyme, gel filtration | Porphyromonas gingivalis |
60880 | - |
truncated PAD with its poly-His and Xpress epitope tags, calculated from amino acid sequence | Porphyromonas gingivalis |
60890 | - |
MALDI-TOF mass spectrometry | Porphyromonas gingivalis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
protein L-Arg + H2O | Porphyromonas gingivalis | - |
protein L-citrulline + NH3 | - |
? | |
protein L-Arg + H2O | Porphyromonas gingivalis BAA-308-5 (W83) | - |
protein L-citrulline + NH3 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Porphyromonas gingivalis | - |
- |
- |
Porphyromonas gingivalis BAA-308-5 (W83) | - |
- |
- |
Purification (Comment) | Organism |
---|---|
Ni+-NTA resin column chromatography, ultrafiltration, and gel filtration | Porphyromonas gingivalis |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.000345 | - |
crude extract, pH 9.5, at 37°C | Porphyromonas gingivalis |
0.175 | - |
after purification, pH 9.5, at 37°C | Porphyromonas gingivalis |
Storage Stability | Organism |
---|---|
25°C or -80°C, in the presence of 0.5 mM 2-mercaptoethanol or 1 mM dithiothreitol and 0.1% (w/v) sodium azide in 50 mM potassium phosphate buffer (pH 7.8), 2 weeks, no appreciable loss of activity | Porphyromonas gingivalis |
4°C or -20°C, in the presence of 0.5 mM 2-mercaptoethanol or 1 mM dithiothreitol and 0.1% (w/v) sodium azide in 50 mM potassium phosphate buffer (pH 7.8), 2 weeks, aggregation of the protein occurs | Porphyromonas gingivalis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
agmatine-L-Arg + H2O | 3.2% activity compared to L-Arg | Porphyromonas gingivalis | N5-carbamoyl-N2-[4-(carbamoylamino)butyl]-L-ornithine + NH3 | - |
? | |
benzoyl-L-arginine ethyl ester + H2O | 8.1% activity compared to L-Arg | Porphyromonas gingivalis | benzoyl-L-citrulline ethyl ester + NH3 | - |
? | |
benzoyl-L-arginine ethyl ester + H2O | 8.1% activity compared to L-Arg | Porphyromonas gingivalis BAA-308-5 (W83) | benzoyl-L-citrulline ethyl ester + NH3 | - |
? | |
bradykinin-L-Arg + H2O | 18.8% activity compared to L-Arg | Porphyromonas gingivalis | bradykinin-L-citrulline + NH3 | - |
? | |
D-Arg + H2O | 26.4% activity compared to L-Arg | Porphyromonas gingivalis | ? | - |
? | |
homoarginine + H2O | 50% activity compared to L-Arg | Porphyromonas gingivalis | (2S)-2-amino-6-[(aminocarbonyl)amino]hexanoic acid + NH3 | - |
? | |
KRPPGFSPL + H2O | 3.1% activity compared to L-Arg | Porphyromonas gingivalis | L-lysyl-N5-carbamoyl-L-ornithyl-L-prolyl-L-prolylglycyl-L-phenylalanyl-L-seryl-L-prolyl-L-leucine + NH3 | - |
? | |
L-Arg + H2O | 100% activity | Porphyromonas gingivalis | citrulline + NH3 | - |
? | |
L-Arg + H2O | 100% activity | Porphyromonas gingivalis BAA-308-5 (W83) | citrulline + NH3 | - |
? | |
L-arginine hydroxamate + H2O | 68.2% activity compared to L-Arg | Porphyromonas gingivalis | ? | - |
? | |
L-canavanine-L-Arg + H2O | 2.5% activity compared to L-Arg | Porphyromonas gingivalis | O-ureido-L-homoserine + NH3 | - |
? | |
L-Pro-L-Arg-L-Phe + H2O | 3.4% activity compared to L-Arg | Porphyromonas gingivalis | ? | - |
? | |
L-Pro-L-Phe-L-Arg + H2O | 78.2% activity compared to L-Arg | Porphyromonas gingivalis | ? | - |
? | |
additional information | no activity with Nomega,Nomega-dimethyl-L-arginine | Porphyromonas gingivalis | ? | - |
? | |
additional information | no activity with Nomega,Nomega-dimethyl-L-arginine | Porphyromonas gingivalis BAA-308-5 (W83) | ? | - |
? | |
N-alpha-benzoyl-L-arginine + H2O | 54.5% activity compared to L-Arg | Porphyromonas gingivalis | N-alpha-benzoyl-L-citrulline + NH3 | - |
? | |
N-alpha-benzoyl-L-arginine + H2O | 54.5% activity compared to L-Arg | Porphyromonas gingivalis BAA-308-5 (W83) | N-alpha-benzoyl-L-citrulline + NH3 | - |
? | |
Nomega-hydroxy-L-arginine + H2O | 2.1% activity compared to L-Arg | Porphyromonas gingivalis | Nomega-hydroxy-L-citrulline + NH3 | - |
? | |
protein L-Arg + H2O | - |
Porphyromonas gingivalis | protein L-citrulline + NH3 | - |
? | |
protein L-Arg + H2O | - |
Porphyromonas gingivalis BAA-308-5 (W83) | protein L-citrulline + NH3 | - |
? |
Synonyms | Comment | Organism |
---|---|---|
PAD | - |
Porphyromonas gingivalis |
peptidylarginine deiminase | - |
Porphyromonas gingivalis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
55 | - |
- |
Porphyromonas gingivalis |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
4 | - |
PAD is cold sensitive, it aggregates within 30 min at 4°C, but aggregated PAD displays variable activity for up to one week after re-suspension with storage buffer | Porphyromonas gingivalis |
100 | - |
no product formation is observed in reaction mixtures containing the PAD sample that have been boiled | Porphyromonas gingivalis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0046 | - |
Nomega-hydroxy-L-arginine | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis | |
0.0052 | - |
L-canavanine-L-Arg | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis | |
0.0068 | - |
bradykinin antagonist-L-Arg | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis | |
0.007 | - |
agmatine-L-Arg | - |
Porphyromonas gingivalis | |
0.0075 | - |
L-Pro-L-Arg-L-Phe | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis | |
0.019 | - |
benzoyl-L-arginine ethyl ester | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis | |
0.043 | - |
bradykinin-L-Arg | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis | |
0.058 | - |
D-Arg | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis | |
0.11 | - |
homoarginine | - |
Porphyromonas gingivalis | |
0.12 | - |
N-alpha-benzoyl-L-arginine | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis | |
0.15 | - |
L-Arginine hydroxamate | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis | |
0.17 | - |
L-Pro-L-Phe-L-Arg | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis | |
0.22 | - |
L-Arg | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
9.5 | - |
- |
Porphyromonas gingivalis |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6 | 11 | 38.5% activity remaining at pH 6.0 and 3.4% activity remaining at pH 11.0 | Porphyromonas gingivalis |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
7 | 9.5 | the enzyme at 13 mg/ml remains active at pH 7.0, 8.0 and 9.5 for several days at 25°C | Porphyromonas gingivalis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | PAD does not need a cofactor for catalysis and stability | Porphyromonas gingivalis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00023 | - |
agmatine-L-Arg | - |
Porphyromonas gingivalis | |
0.00093 | - |
L-canavanine-L-Arg | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis | |
0.00099 | - |
benzoyl-L-arginine ethyl ester | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis | |
0.003 | - |
Nomega-hydroxy-L-arginine | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis | |
0.022 | - |
L-Arginine hydroxamate | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis | |
0.024 | - |
homoarginine | - |
Porphyromonas gingivalis | |
0.031 | - |
D-Arg | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis | |
0.11 | - |
L-Pro-L-Arg-L-Phe | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis | |
0.19 | - |
L-Arg | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis | |
0.21 | - |
bradykinin antagonist-L-Arg | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis | |
1.9 | - |
L-Pro-L-Phe-L-Arg | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis | |
2.1 | - |
bradykinin-L-Arg | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis | |
6 | 12 | N-alpha-benzoyl-L-arginine | in 50 mM CHES/HCl, pH 9.5 containing 10 mM dithiothreitol, at 37°C | Porphyromonas gingivalis |