EC Number | Cloned (Comment) | Organism |
---|---|---|
1.8.4.9 | expression of N-terminally His-tagged PpAPR in Escherichia coli, expression of GFP-tagged APR in Physcomitrella patens plants using the mAV4 vector containing the chloroplast transit peptide from Physcomitrella patens FtsZ2-1 and transient transfection of protoplasts | Physcomitrium patens |
1.8.4.9 | overexpression in Escherichia coli | Physcomitrium patens |
EC Number | Inhibitors | Comment | Organism | Structure |
---|---|---|---|---|
1.8.4.9 | adenosine 5'-phosphosulfate | inhibits activity with 3'-phosphoadenosine 5'-phosphosulfate | Physcomitrium patens | |
1.8.4.9 | glutathione | inhibits APR at concentrations above 30 mM | Physcomitrium patens | |
1.8.4.9 | KNO3 | abolishes enzyme activation by other salts and inhibits the enzyme | Physcomitrium patens |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.8.4.9 | 0.02 | - |
3'-phosphoadenosine 5'-phosphosulfate | - |
Physcomitrium patens | |
1.8.4.9 | 0.05 | - |
adenosine 5'-phosphosulfate | - |
Physcomitrium patens |
EC Number | Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|---|
1.8.4.9 | chloroplast | - |
Physcomitrium patens | 9507 | - |
1.8.4.9 | chloroplast | determination of subcellular localization, overview | Physcomitrium patens | 9507 | - |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
1.8.4.9 | (NH4)2SO4 | highly activating | Physcomitrium patens | |
1.8.4.9 | (NH4)2SO4 | 500 mM, stimulates | Physcomitrium patens | |
1.8.4.9 | Fe2+ | FeS cluster as a cofactor in PpAPR | Physcomitrium patens | |
1.8.4.9 | KNO3 | 500 mM, stimulates | Physcomitrium patens | |
1.8.4.9 | Mg2+ | activates 18-24fold at 500 mM | Physcomitrium patens | |
1.8.4.9 | MgCl2 | 500 mM, stimulates | Physcomitrium patens | |
1.8.4.9 | MgSO4 | 500 mM, stimulates | Physcomitrium patens | |
1.8.4.9 | additional information | no iron is associated with recombinant PpPAPR | Physcomitrium patens | |
1.8.4.10 | (NH4)2SO4 | activates | Physcomitrium patens | |
1.8.4.10 | KNO3 | activates | Physcomitrium patens | |
1.8.4.10 | Mg2+ | activates | Physcomitrium patens | |
1.8.4.10 | additional information | PpAPR-B does not contain the FeS cluster, which is believed to determine the substrate specificity of other APR enzymes from seed plants. The lack of the FeS cluster in PpAPR-B catalysis is connected with a lower turnover rate but higher stability of the protein | Physcomitrium patens | |
1.8.4.10 | sulfate | activates | Physcomitrium patens |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
1.8.4.9 | 32000 | - |
x * 32000, SDS-PAGE | Physcomitrium patens |
1.8.4.9 | 50000 | - |
x * 50000, recombinant His-tagged PpAPR, SDS-PAGE | Physcomitrium patens |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.8.4.9 | adenylyl sulfate + glutathione | Physcomitrium patens | sulfate assimilation pathway in Physcomitrella patens, overview | AMP + sulfite + glutathione disulfide | - |
? | |
1.8.4.9 | additional information | Physcomitrium patens | the moss Physcomitrella patens is unique among these organisms in possessing orthologs of both APR and PAPR, EC 1.8.4.8, genes | ? | - |
? | |
1.8.4.10 | adenylyl sulfate + thioredoxin | Physcomitrium patens | sulfate assimilation pathway in Physcomitrella patens, overview | AMP + sulfite + thioredoxin disulfide | - |
? | |
1.8.4.10 | additional information | Physcomitrium patens | the moss Physcomitrella patens is unique among these organisms in possessing orthologs of both APR and PAPR, EC 1.8.4.8, genes | ? | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.8.4.9 | Physcomitrium patens | - |
- |
- |
1.8.4.10 | Physcomitrium patens | - |
- |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
1.8.4.9 | recombinant N-terminally His-tagged PpAPR from Escherichia coli | Physcomitrium patens |
EC Number | Reaction | Comment | Organism | Reaction ID |
---|---|---|---|---|
1.8.4.9 | AMP + sulfite + glutathione disulfide = adenylyl sulfate + 2 glutathione | the reaction catalyzed by APR can be divided into three steps. In the first step, APS binds to the protein, and a reductive transfer results in sulfite bound to the active-site cysteine in a stable reaction intermediate. In the second step, free sulfite is released by the action of the C-terminal domain. In the third step, the glutaredoxin active site is regenerated by reduction with GSH reductase | Physcomitrium patens | |
1.8.4.10 | AMP + sulfite + thioredoxin disulfide = 5'-adenylyl sulfate + thioredoxin | the reaction catalyzed by APR can be divided into three steps. In the first step, APS binds to the protein, and a reductive transfer results in sulfite bound to the active-site cysteine in a stable reaction intermediate. In the second step, free sulfite is released by the action of free thioredoxin. In the third step, the thioredoxin is regenerated by reduction with thioredoxin reductase | Physcomitrium patens |
EC Number | Storage Stability | Organism |
---|---|---|
1.8.4.9 | -20°C, 1 day, PpAPR is completely inactivated, plant APRs are very unstable enzymes because of the rapid loss of their iron-sulfur clusters | Physcomitrium patens |
1.8.4.9 | -20°C, 1 day, PpAPR-B retains 30% of its activity | Physcomitrium patens |
1.8.4.9 | 4°C, PpAPR, completely stable for 48 h, inactivation after longer than 2 dayswith 15% remaining activity after 4-5 days | Physcomitrium patens |
1.8.4.9 | 4°C, PpAPR-B is stable over 5 days | Physcomitrium patens |
1.8.4.10 | -20°C, 1 day, PpAPR-B retains 30% of its activity | Physcomitrium patens |
1.8.4.10 | 4°C, PpAPR-B, completely stable for 5 days | Physcomitrium patens |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.8.4.9 | 3'-phosphoadenosine 5'-phosphosulfate + dithioerythritol | - |
Physcomitrium patens | ? | - |
? | |
1.8.4.9 | 3'-phosphoadenosine 5'-phosphosulfate + glutathione | - |
Physcomitrium patens | adenosine 3',5'-diphosphate + sulfite + oxidized glutathione | - |
? | |
1.8.4.9 | adenosine 5'-phosphosulfate + glutathione | PpAPR-B is much more active with adenosine 5'-phosphosulfate as a substrate than with 3'-phosphoadenosine 5'-phosphosulfate | Physcomitrium patens | ? | - |
? | |
1.8.4.9 | adenylyl sulfate + DTT | - |
Physcomitrium patens | ? | - |
? | |
1.8.4.9 | adenylyl sulfate + glutathione | sulfate assimilation pathway in Physcomitrella patens, overview | Physcomitrium patens | AMP + sulfite + glutathione disulfide | - |
? | |
1.8.4.9 | adenylyl sulfate + glutathione | substrate binding structure, modelling, overview | Physcomitrium patens | AMP + sulfite + glutathione disulfide | - |
? | |
1.8.4.9 | additional information | no activity with thioredoxin | Physcomitrium patens | ? | - |
? | |
1.8.4.9 | additional information | the moss Physcomitrella patens is unique among these organisms in possessing orthologs of both APR and PAPR, EC 1.8.4.8, genes | Physcomitrium patens | ? | - |
? | |
1.8.4.10 | adenylyl sulfate + thioredoxin | sulfate assimilation pathway in Physcomitrella patens, overview | Physcomitrium patens | AMP + sulfite + thioredoxin disulfide | - |
? | |
1.8.4.10 | adenylyl sulfate + thioredoxin | dependent on thioredoxin, APS is the preferred substrate, substrate binding structure, modelling, overview | Physcomitrium patens | AMP + sulfite + thioredoxin disulfide | - |
? | |
1.8.4.10 | additional information | the moss Physcomitrella patens is unique among these organisms in possessing orthologs of both APR and PAPR, EC 1.8.4.8, genes | Physcomitrium patens | ? | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
1.8.4.9 | ? | x * 32000, SDS-PAGE | Physcomitrium patens |
1.8.4.9 | ? | x * 50000, recombinant His-tagged PpAPR, SDS-PAGE | Physcomitrium patens |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.8.4.9 | APR | - |
Physcomitrium patens |
1.8.4.9 | PpAPR-B | - |
Physcomitrium patens |
1.8.4.10 | APR | - |
Physcomitrium patens |
1.8.4.10 | PpAPR-B | - |
Physcomitrium patens |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
1.8.4.9 | 37 | - |
- |
Physcomitrium patens |
1.8.4.10 | 37 | - |
- |
Physcomitrium patens |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.8.4.9 | 0.00075 | - |
3'-phosphoadenosine 5'-phosphosulfate | - |
Physcomitrium patens | |
1.8.4.9 | 0.176 | - |
adenosine 5'-phosphosulfate | - |
Physcomitrium patens |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
1.8.4.9 | 7 | 8.5 | - |
Physcomitrium patens |
1.8.4.9 | 8 | - |
substrate: 3'-phosphoadenosine 5'-phosphosulfate | Physcomitrium patens |
1.8.4.10 | additional information | - |
PpAPR-B activity in terms of APS reduction increases gradually and less markedly with increasing pH compared to PpAPR, EC 1.8.4.9 | Physcomitrium patens |
1.8.4.10 | 8 | - |
- |
Physcomitrium patens |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
1.8.4.9 | glutathione | - |
Physcomitrium patens | |
1.8.4.9 | additional information | FeS cluster as a cofactor in PpAPR, no activity with thioredoxin | Physcomitrium patens | |
1.8.4.10 | additional information | PpAPR-B does not contain the FeS cluster, which is believed to determine the substrate specificity of other APR enzymes from seed plants. The lack of the FeS cluster in PpAPR-B catalysis is connected with a lower turnover rate but higher stability of the protein. No activity with glutathione or DTT | Physcomitrium patens | |
1.8.4.10 | thioredoxin | - |
Physcomitrium patens |
EC Number | Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.8.4.9 | 0.176 | - |
adenosine 5'-phosphosulfate | - |
Physcomitrium patens |