Application | Comment | Organism |
---|---|---|
medicine | tobacco PPO2 represents a useful model system for the understanding of the structurefunction relationship underlying detrimental human enzyme defects, Arg98, Phe392, Leu356 and Leu372 are functionally involved in substrate co-ordination within the active site | Nicotiana tabacum |
Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli strain BL21(DE3)RIL | Nicotiana tabacum |
expression in Escherichia coli BL21(DE3)RIL containing the original or mutated pET32a-PPO2 plasmids | Nicotiana tabacum |
Crystallization (Comment) | Organism |
---|---|
- |
Nicotiana tabacum |
Protein Variants | Comment | Organism |
---|---|---|
F329E | shows increased activity | Nicotiana tabacum |
F392E | inferior binding of the substrate, but increased turnover rate than wild-type | Nicotiana tabacum |
F392H | does not reveal detectable enzyme activity indicating an important role of Phe392 in substrate ring A stacking | Nicotiana tabacum |
F392H | no detectable enzyme activity | Nicotiana tabacum |
L356N | an only slightly inferior binding capacity compared with the wild-type and comparable catalytic PPO2 activities | Nicotiana tabacum |
L356N | shows increased activity | Nicotiana tabacum |
L356V | increased Km value, once the substrate is bound, catalysis is enhanced | Nicotiana tabacum |
L356V | shows increased activity | Nicotiana tabacum |
L372N | an only slightly inferior binding capacity compared with the wild-type and comparable catalytic PPO2 activities | Nicotiana tabacum |
L372N | shows increased activity | Nicotiana tabacum |
L372V | increased Km value, once the substrate is bound, catalysis is enhanced | Nicotiana tabacum |
L372V | shows increased activity | Nicotiana tabacum |
N67R | inferior substrate binding, acceptable catalytic efficiency | Nicotiana tabacum |
N67R | shows increased activity | Nicotiana tabacum |
N67R/S374D | almost wild-type level Km value | Nicotiana tabacum |
N67R/S374D | shows increased activity | Nicotiana tabacum |
N67W | does not reveal detectable enzyme activity | Nicotiana tabacum |
N67W/S374D | significantly reduced enzyme activity | Nicotiana tabacum |
N67W/S374D | shows increased activity, structural constellation mimicking the human disease variegate porphyria | Nicotiana tabacum |
N6W | no detectable enzyme activity | Nicotiana tabacum |
R98A | 8fold increased Km value, improved turnover | Nicotiana tabacum |
R98A | shows increased activity | Nicotiana tabacum |
R98E | inferior substrate binding, improved turnover | Nicotiana tabacum |
R98E | shows increased activity | Nicotiana tabacum |
R98K | improved turnover | Nicotiana tabacum |
R98K | shows increased activity | Nicotiana tabacum |
S374D | increased turnover rate | Nicotiana tabacum |
S374D | shows increased activity | Nicotiana tabacum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00117 | - |
protoporphyrinogen-IX | wild-type | Nicotiana tabacum | |
0.00117 | - |
protoporphyrinogen-IX | wild type enzyme PPO2, at 20°C | Nicotiana tabacum | |
0.0014 | - |
protoporphyrinogen-IX | N67R/S374D | Nicotiana tabacum | |
0.0014 | - |
protoporphyrinogen-IX | mutant enzyme N67R/S374D, at 20°C | Nicotiana tabacum | |
0.0026 | - |
protoporphyrinogen-IX | R98K | Nicotiana tabacum | |
0.0026 | - |
protoporphyrinogen-IX | mutant enzyme R98K, at 20°C | Nicotiana tabacum | |
0.0073 | - |
protoporphyrinogen-IX | L356V | Nicotiana tabacum | |
0.0073 | - |
protoporphyrinogen-IX | mutant enzyme L356V, at 20°C | Nicotiana tabacum | |
0.0083 | - |
protoporphyrinogen-IX | R98A | Nicotiana tabacum | |
0.0083 | - |
protoporphyrinogen-IX | mutant enzyme R98A, at 20°C | Nicotiana tabacum | |
0.0109 | - |
protoporphyrinogen-IX | S374D | Nicotiana tabacum | |
0.0109 | - |
protoporphyrinogen-IX | mutant enzyme S374D, at 20°C | Nicotiana tabacum | |
0.011 | - |
protoporphyrinogen-IX | L356N | Nicotiana tabacum | |
0.011 | - |
protoporphyrinogen-IX | mutant enzyme L356N, at 20°C | Nicotiana tabacum | |
0.0112 | - |
protoporphyrinogen-IX | F392E | Nicotiana tabacum | |
0.0112 | - |
protoporphyrinogen-IX | mutant enzyme F392E, at 20°C | Nicotiana tabacum | |
0.0125 | - |
protoporphyrinogen-IX | R98E | Nicotiana tabacum | |
0.0125 | - |
protoporphyrinogen-IX | mutant enzyme R98E, at 20°C | Nicotiana tabacum | |
0.0164 | - |
protoporphyrinogen-IX | L372N | Nicotiana tabacum | |
0.0164 | - |
protoporphyrinogen-IX | mutant enzyme L372N, at 20°C | Nicotiana tabacum | |
0.021 | - |
protoporphyrinogen-IX | N67W/S374D | Nicotiana tabacum | |
0.021 | - |
protoporphyrinogen-IX | mutant enzyme N67W/S374D, at 20°C | Nicotiana tabacum | |
0.097 | - |
protoporphyrinogen-IX | N67R | Nicotiana tabacum | |
0.097 | - |
protoporphyrinogen-IX | mutant enzyme N67R, at 20°C | Nicotiana tabacum | |
0.103 | - |
protoporphyrinogen-IX | L372V | Nicotiana tabacum | |
0.103 | - |
protoporphyrinogen-IX | mutant enzyme L372V, at 20°C | Nicotiana tabacum |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
mitochondrion | PPO2 | Nicotiana tabacum | 5739 | - |
plastid | PPO1 | Nicotiana tabacum | 9536 | - |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Nicotiana tabacum | - |
- |
- |
Purification (Comment) | Organism |
---|---|
Ni-IDA resin column chromatography | Nicotiana tabacum |
to apparent homogeneity by Ni-IDA resin | Nicotiana tabacum |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
4.27 | - |
- |
Nicotiana tabacum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
protoporphyrinogen-IX + 3 O2 | - |
Nicotiana tabacum | protoporphyrin-IX + 3 H2O2 | - |
? | |
protoporphyrinogen-IX + 3 O2 | - |
Nicotiana tabacum | protoporphyrin-IX + 3 H2O2 | - |
ir |
Synonyms | Comment | Organism |
---|---|---|
PPO1 | plastidic PPO | Nicotiana tabacum |
PPO2 | - |
Nicotiana tabacum |
PPO2 | mitochondrial PPO | Nicotiana tabacum |
protoporphyrinogen IX oxidase | - |
Nicotiana tabacum |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
6 | - |
protoporphyrinogen-IX | wild-type | Nicotiana tabacum | |
6 | - |
protoporphyrinogen-IX | wild type enzyme PPO2, at 20°C | Nicotiana tabacum | |
7 | - |
protoporphyrinogen-IX | L372N | Nicotiana tabacum | |
7 | - |
protoporphyrinogen-IX | mutant enzyme L372N, at 20°C | Nicotiana tabacum | |
11 | - |
protoporphyrinogen-IX | F392E | Nicotiana tabacum | |
11 | - |
protoporphyrinogen-IX | mutant enzyme F392E, at 20°C | Nicotiana tabacum | |
12 | - |
protoporphyrinogen-IX | N67W/S374D | Nicotiana tabacum | |
12 | - |
protoporphyrinogen-IX | mutant enzyme N67W/S374D, at 20°C | Nicotiana tabacum | |
34 | - |
protoporphyrinogen-IX | R98E | Nicotiana tabacum | |
34 | - |
protoporphyrinogen-IX | mutant enzyme R98E, at 20°C | Nicotiana tabacum | |
37 | - |
protoporphyrinogen-IX | R98K | Nicotiana tabacum | |
37 | - |
protoporphyrinogen-IX | mutant enzyme R98K, at 20°C | Nicotiana tabacum | |
38 | - |
protoporphyrinogen-IX | L356N | Nicotiana tabacum | |
38 | - |
protoporphyrinogen-IX | mutant enzyme L356N, at 20°C | Nicotiana tabacum | |
97 | - |
protoporphyrinogen-IX | N67R | Nicotiana tabacum | |
97 | - |
protoporphyrinogen-IX | mutant enzyme N67R, at 20°C | Nicotiana tabacum | |
111 | - |
protoporphyrinogen-IX | N67R/S374D | Nicotiana tabacum | |
111 | - |
protoporphyrinogen-IX | mutant enzyme N67R/S374D, at 20°C | Nicotiana tabacum | |
208 | - |
protoporphyrinogen-IX | mutant enzyme S374D, at 20°C | Nicotiana tabacum | |
300 | - |
protoporphyrinogen-IX | L356V | Nicotiana tabacum | |
300 | - |
protoporphyrinogen-IX | mutant enzyme L356V, at 20°C | Nicotiana tabacum | |
365 | - |
protoporphyrinogen-IX | R98A | Nicotiana tabacum | |
365 | - |
protoporphyrinogen-IX | mutant enzyme R98A, at 20°C | Nicotiana tabacum | |
597 | - |
protoporphyrinogen-IX | L372V | Nicotiana tabacum | |
597 | - |
protoporphyrinogen-IX | mutant enzyme L372V, at 20°C | Nicotiana tabacum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
flavin | dependent | Nicotiana tabacum |