Cloned (Comment) | Organism |
---|---|
functional expression of N-terminally His6-tagged enzyme in Escherichia coli strain pLysS (DE3) | Geobacillus stearothermophilus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
aza-S-adenosyl-L-methionine | complete inhibition | Geobacillus stearothermophilus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | iron-sulfur protein, the aerobically purified apo-SplG forms a homodimer, which contains one [4Fe-4S] cluster per monomer unit after reconstitution to the holoform | Geobacillus stearothermophilus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
5,6-dihydro-5-(thymidin-7-yl)thymidine (in double helical DNA) | Geobacillus stearothermophilus | - |
thymidylyl-(3'-5')-thymidylate (in DNA) | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Geobacillus stearothermophilus | - |
gene splG | - |
Purification (Comment) | Organism |
---|---|
recombinant His6-tagged enzyme from Escherichia coli strain pLysS (DE3) by nickel affinity chromatography and gel filtration | Geobacillus stearothermophilus |
Renatured (Comment) | Organism |
---|---|
for reconstitution of the holoenzyme with [4Fe-4S]-cluster, the recombinant apo-SplG is dissolved in 0.8 ml of reconstitution buffer containing 50 mM Tris-HCl, pH 8.0, and 5mM DTT, for 0.5 h, followed by the addition of FeCl2 and Na2S to a final concentration of 100M. The SplG is then incubated anaerobically for 16 h at 4 °C | Geobacillus stearothermophilus |
Source Tissue | Comment | Organism | Textmining |
---|
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
2.6 | - |
purified recombinant enzyme, pH and temperature not specified in the publication | Geobacillus stearothermophilus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(5R)-5,6-dihydro-5-(thymidin-7-yl)thymidine (in double helical DNA) + S-adenosyl-L-methionine + 2 H+ | assay with a synthetic dinucleotide SP lesion substrate and with smallDNAsingle strands, which contain one SP lesion at a defined site | Geobacillus stearothermophilus | thymidylyl-(3'-5')-thymidylate (in DNA) + 5'-deoxyadenosine + L-methionine | - |
? | |
(5S)-5,6-dihydro-5-(thymidin-7-yl)thymidine (in double helical DNA) | assay with a synthetic dinucleotide SP lesion substrate and with smallDNAsingle strands, which contain one SP lesion at a defined site | Geobacillus stearothermophilus | thymidylyl-(3'-5')-thymidylate (in DNA) | - |
? | |
5,6-dihydro-5-(thymidin-7-yl)thymidine (in double helical DNA) | - |
Geobacillus stearothermophilus | thymidylyl-(3'-5')-thymidylate (in DNA) | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | the aerobically purified recombinant apo-SplG forms a homodimer, which contains one [4Fe-4S] cluster permonomerunit after reconstitution to the holoform, secondary SplG structure, overview | Geobacillus stearothermophilus |
Synonyms | Comment | Organism |
---|---|---|
More | the thermophilic spore photoproduct lyase belongs to the family of radical S-adenosylmethionine enzymes | Geobacillus stearothermophilus |
SplG | - |
Geobacillus stearothermophilus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
S-adenosyl-L-methionine | - |
Geobacillus stearothermophilus |