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Literature summary extracted from

  • Kim, S.K.; Gomes, V.; Gao, Y.; Chandramouli, K.; Johnson, M.K.; Knaff, D.B.; Leustek, T.
    The two-domain structure of 5-adenylylsulfate (APS) reductase from Enteromorpha intestinalis is a requirement for efficient APS reductase activity (2007), Biochemistry, 46, 591-601.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.8.99.2 Na2SO4 500 mM, 5 fold stimulation Ulva intestinalis

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.8.4.9 a heterologous system is constructed in which the C domain of EiAPR was fused to the carboxyl terminus of the APS reductase from Pseudomonas aeruginosa (PaAPR), an enzyme that normally uses thioredoxin as an electron donor and is incapable of using glutathione for this function. The hybrid enzyme, which retains the [4Fe-4S] cluster from PaAPR can use both thioredoxin and glutathione as an electron donor for 5'-adenylylsulfate reduction. Expression in Escherichia coli Ulva intestinalis
1.8.99.2 expressed in Escherichia coli Ulva intestinalis

Protein Variants

EC Number Protein Variants Comment Organism
1.8.4.10 additional information a heterologous system is constructed in which the C domain of EiAPR (EC 1.8.4.9) is fused to the carboxyl terminus of the APS reductase from Pseudomonas aeruginosa (EC 1.8.4.10), an enzyme that normally uses thioredoxin as an electron donor and is incapable of using glutathione for this function. The hybrid enzyme, which retains the [4Fe-4S] cluster from PaAPR, can use both thioredoxin and glutathione as an electron donor for APS reduction. The ability to use glutathione is enhanced by the addition of Na2SO4 to the reaction buffer, a property that the hybrid enzyme shares with EiAPR Pseudomonas aeruginosa

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.8.4.9 0.0002
-
 glutathione
-
 Ulva intestinalis
1.8.4.9 0.011
-
 5'-adenylyl sulfate
-
 Ulva intestinalis
1.8.4.10 0.0114
-
 5'-adenylyl sulfate
-
 Pseudomonas aeruginosa
1.8.4.10 0.054
-
 thioredoxin
-
 Pseudomonas aeruginosa
1.8.99.2 0.0002
-
 dithiothreitol
-
 Ulva intestinalis
1.8.99.2 0.011
-
 5'-adenylylsulfate
-
 Ulva intestinalis

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.8.4.10 Fe-S cluster [4Fe-4S] cluster Pseudomonas aeruginosa

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.8.99.2 20000
-
 calculated from sequence of cDNA, C-terminal domain of EiAPR Ulva intestinalis
1.8.99.2 20000
-
 MALDI-TOF mass spectrometry, C-terminal domain of EiAPR Ulva intestinalis
1.8.99.2 48700
-
 MALDI-TOF mass spectrometry, whole enzyme Ulva intestinalis
1.8.99.2 48710
-
 calculated from sequence of cDNA, whole enzyme Ulva intestinalis

Organism

EC Number Organism UniProt Comment Textmining
1.8.4.9 Ulva intestinalis
-
-
-
1.8.4.10 Pseudomonas aeruginosa
-
-
-
1.8.99.2 Ulva intestinalis
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.8.4.9
-
 Ulva intestinalis
1.8.99.2 Ni2+-affinity column chromatography Ulva intestinalis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.8.4.9 5'-adenylyl sulfate + glutathione
-
 Ulva intestinalis AMP + sulfite + glutathione disulfide
-
 ?
1.8.4.10 5'-adenylyl sulfate + thioredoxin
-
 Pseudomonas aeruginosa AMP + sulfite + thioredoxin disulfide
-
 ?
1.8.99.2 5'-adenylylsulfate + dithiothreitol
-
 Ulva intestinalis adenosine 5'-monophosphate + sulfite + oxidized dithiothreitol
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.8.4.9 EiAPR
-
 Ulva intestinalis
1.8.4.10 PaAPR
-
 Pseudomonas aeruginosa
1.8.99.2 5-adenylylsulfate reductase
-
 Ulva intestinalis
1.8.99.2 EiAPR
-
 Ulva intestinalis

pI Value

EC Number Organism Comment pI Value Maximum pI Value
1.8.99.2 Ulva intestinalis
-
-
 6.91