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Literature summary for 1.8.4.9 extracted from

  • Kopriva, S.; Fritzemeier, K.; Wiedemann, G.; Reski, R.
    The putative moss 3'-phosphoadenosine-5'-phosphosulfate reductase is a novel form of adenosine-5-phosphosulfate reductase without an iron-sulfur cluster (2007), J. Biol. Chem., 282, 22930-22938.
    View publication on PubMed

Cloned(Commentary)

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

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 [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

Localization Comment Organism GeneOntology No. Textmining
chloroplast
-
Physcomitrium patens 9507
-
chloroplast determination of subcellular localization, overview Physcomitrium patens 9507
-

Metals/Ions

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 [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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Physcomitrium patens
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant N-terminally His-tagged PpAPR from Escherichia coli Physcomitrium patens

Reaction

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

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 and Products (Substrate)

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

Subunits Comment Organism
? x * 32000, SDS-PAGE Physcomitrium patens
? x * 50000, recombinant His-tagged PpAPR, SDS-PAGE Physcomitrium patens

Synonyms

Synonyms Comment Organism
APR
-
Physcomitrium patens
PpAPR-B
-
Physcomitrium patens

Temperature Optimum [┬░C]

Temperature Optimum [┬░C] Temperature Optimum Maximum [┬░C] Comment Organism
37
-
-
Physcomitrium patens

Turnover Number [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
7 8.5
-
Physcomitrium patens
8
-
substrate: 3'-phosphoadenosine 5'-phosphosulfate Physcomitrium patens

Cofactor

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 [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.176
-
adenosine 5'-phosphosulfate
-
Physcomitrium patens