Cloned (Comment) | Organism |
---|---|
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 |
overexpression in Escherichia coli | Physcomitrium patens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
adenosine 5'-phosphosulfate | inhibits activity with 3'-phosphoadenosine 5'-phosphosulfate | Physcomitrium patens | |
glutathione | inhibits APR at concentrations above 30 mM | Physcomitrium patens | |
KNO3 | abolishes enzyme activation by other salts and inhibits the enzyme | Physcomitrium patens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.02 | - |
3'-phosphoadenosine 5'-phosphosulfate | - |
Physcomitrium patens | |
0.05 | - |
adenosine 5'-phosphosulfate | - |
Physcomitrium patens |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
chloroplast | - |
Physcomitrium patens | 9507 | - |
chloroplast | determination of subcellular localization, overview | Physcomitrium patens | 9507 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
(NH4)2SO4 | highly activating | Physcomitrium patens | |
(NH4)2SO4 | 500 mM, stimulates | Physcomitrium patens | |
Fe2+ | FeS cluster as a cofactor in PpAPR | Physcomitrium patens | |
KNO3 | 500 mM, stimulates | Physcomitrium patens | |
Mg2+ | activates 18-24fold at 500 mM | Physcomitrium patens | |
MgCl2 | 500 mM, stimulates | Physcomitrium patens | |
MgSO4 | 500 mM, stimulates | Physcomitrium patens | |
additional information | no iron is associated with recombinant PpPAPR | Physcomitrium patens |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
32000 | - |
x * 32000, SDS-PAGE | Physcomitrium patens |
50000 | - |
x * 50000, recombinant His-tagged PpAPR, SDS-PAGE | Physcomitrium patens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
adenylyl sulfate + glutathione | Physcomitrium patens | sulfate assimilation pathway in Physcomitrella patens, overview | AMP + sulfite + glutathione disulfide | - |
? | |
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 | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Physcomitrium patens | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant N-terminally His-tagged PpAPR from Escherichia coli | Physcomitrium patens |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
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 |
Storage Stability | Organism |
---|---|
-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 |
-20°C, 1 day, PpAPR-B retains 30% of its activity | Physcomitrium patens |
4°C, PpAPR, completely stable for 48 h, inactivation after longer than 2 dayswith 15% remaining activity after 4-5 days | Physcomitrium patens |
4°C, PpAPR-B is stable over 5 days | Physcomitrium patens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
3'-phosphoadenosine 5'-phosphosulfate + dithioerythritol | - |
Physcomitrium patens | ? | - |
? | |
3'-phosphoadenosine 5'-phosphosulfate + glutathione | - |
Physcomitrium patens | adenosine 3',5'-diphosphate + sulfite + oxidized glutathione | - |
? | |
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 | ? | - |
? | |
adenylyl sulfate + DTT | - |
Physcomitrium patens | ? | - |
? | |
adenylyl sulfate + glutathione | sulfate assimilation pathway in Physcomitrella patens, overview | Physcomitrium patens | AMP + sulfite + glutathione disulfide | - |
? | |
adenylyl sulfate + glutathione | substrate binding structure, modelling, overview | Physcomitrium patens | AMP + sulfite + glutathione disulfide | - |
? | |
additional information | no activity with thioredoxin | Physcomitrium patens | ? | - |
? | |
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 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 32000, SDS-PAGE | Physcomitrium patens |
? | x * 50000, recombinant His-tagged PpAPR, SDS-PAGE | Physcomitrium patens |
Synonyms | Comment | Organism |
---|---|---|
APR | - |
Physcomitrium patens |
PpAPR-B | - |
Physcomitrium patens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
- |
Physcomitrium patens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00075 | - |
3'-phosphoadenosine 5'-phosphosulfate | - |
Physcomitrium patens | |
0.176 | - |
adenosine 5'-phosphosulfate | - |
Physcomitrium patens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | 8.5 | - |
Physcomitrium patens |
8 | - |
substrate: 3'-phosphoadenosine 5'-phosphosulfate | Physcomitrium patens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
glutathione | - |
Physcomitrium patens | |
additional information | FeS cluster as a cofactor in PpAPR, no activity with thioredoxin | Physcomitrium patens |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.176 | - |
adenosine 5'-phosphosulfate | - |
Physcomitrium patens |