Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli M15(pREP4) cells | Pseudomonas aeruginosa |
gene cpdA, DNA and mino acid sequenc determination and analysis, expression of His-tagged enzyme in Escherichia coli strain M15(pREP4) | Pseudomonas aeruginosa |
Protein Variants | Comment | Organism |
---|---|---|
D63A | the mutant shows less than 0.1% of wild type CpdA activity | Pseudomonas aeruginosa |
H23A | the mutant shows less than 0.1% of wild type CpdA activity | Pseudomonas aeruginosa |
N93A | the mutant shows less than 0.1% of wild type CpdA activity | Pseudomonas aeruginosa |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
alpha-alpha'-dipyridyl | treatment of CpdA with the Fe2+-specific chelator alpha-alpha'-dipyridyl results in a nearly complete loss of activity | Pseudomonas aeruginosa |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0067 | - |
3',5'-cAMP | the addition of FeCl2 does not significantly influence substrate affinity of CdpA increases the rate of the 5'-AMP production, pH and temperature not specified in the publication | Pseudomonas aeruginosa |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | Fe2+ is required for enzyme activity. The catalytic mechanism for CpdA utilizes a Fe3+-Fe2+ center. Addition of 0.01 mM FeCl2, as a source of Fe2+, results in an 2fold stimulation of CpdA activity | Pseudomonas aeruginosa | |
Fe2+ | iron and conserved residues are essential for CpdA activity | Pseudomonas aeruginosa | |
additional information | the addition of Mg2+, Mn2+, Zn2+, and Ca2+ has no effect on CdpA | Pseudomonas aeruginosa | |
additional information | no effect on enzyme activity by Mg2+, Mn2+, Zn2+, and Ca2+ | Pseudomonas aeruginosa |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
31000 | - |
1 * 31000 Da, SDS-PAGE | Pseudomonas aeruginosa |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
3',5'-cAMP + H2O | Pseudomonas aeruginosa | CpdA possesses 3',5'-cAMP phosphodiesterase activity in vitro | 5'-AMP | - |
? | |
additional information | Pseudomonas aeruginosa | iron and conserved residues are essential for CpdA activity, the catalytic mechanism for Pseudomonas aeruginosa CpdA utilizes a Fe3+-Fe2+ center | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas aeruginosa | D4P095 | gene cpdA | - |
Pseudomonas aeruginosa | D4P095 | strain PAK | - |
Purification (Comment) | Organism |
---|---|
Ni-NTA column chromatography and Sephacryl S-200 gel filtration | Pseudomonas aeruginosa |
recombinant His-tagged enzyme from Escherichia coli strain M15(pREP4) by nickel affinity chromatography | Pseudomonas aeruginosa |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
3',5'-cAMP + H2O | CpdA possesses 3',5'-cAMP phosphodiesterase activity in vitro | Pseudomonas aeruginosa | 5'-AMP | - |
? | |
adenosine 3',5'-cyclic phosphate + H2O | - |
Pseudomonas aeruginosa | adenosine 5'-phosphate | - |
? | |
additional information | iron and conserved residues are essential for CpdA activity, the catalytic mechanism for Pseudomonas aeruginosa CpdA utilizes a Fe3+-Fe2+ center | Pseudomonas aeruginosa | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
monomer | 1 * 31000 Da, SDS-PAGE | Pseudomonas aeruginosa |
Synonyms | Comment | Organism |
---|---|---|
cAMP phosphodiesterase | - |
Pseudomonas aeruginosa |
CpdA | - |
Pseudomonas aeruginosa |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Pseudomonas aeruginosa |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.6 | - |
assay at | Pseudomonas aeruginosa |
Organism | Comment | Expression |
---|---|---|
Pseudomonas aeruginosa | the cAMP-dependent transcription factor Vfr directly activates expression of cpdA in response to elevated intracellular cAMP | up |
Pseudomonas aeruginosa | cpdA expression is positively regulated by cAMP-Vfr. cAMP-Vfr binds to the cpdA promoter region, suggesting that in vivo, cpdA transcription is directly activated by cAMP-Vfr | up |
General Information | Comment | Organism |
---|---|---|
malfunction | deletion of cpdA results in the accumulation of intracellular cAMP and altered regulation of Pseudomonas aeruginosa virulence traits | Pseudomonas aeruginosa |
malfunction | deletion of cpdA results in the accumulation of intracellular cAMP and altered regulation of Pseudomonas aeruginosa virulence traits. The cpdA mutant has a cAMP-independent small-colony, slow-growth phenotype | Pseudomonas aeruginosa |
physiological function | CpdA is required for cAMP homeostasis and virulence factor regulation, CpdA affects vfr expression and Vfr protein levels and production of virulence factors ExoS, ToxA, and protease IV | Pseudomonas aeruginosa |
physiological function | CpdA possesses 3',5'-cAMP phosphodiesterase activity in vitro and that it utilizes an iron-dependent catalytic mechanism. The cAMP-dependent transcription factor Vfr directly regulates cpdA expression in response to intracellular cAMP accumulation, thus providing a feedback mechanism for controlling cAMP levels and fine-tuning virulence factor expression. CpdA affects vfr expression and Vfr protein levels. CpdA affects production of virulence factors ExoS, ToxA, and protease IV | Pseudomonas aeruginosa |