Literature summary for 4.1.99.14 extracted from

Pieck, J.C.; Hennecke, U.; Pierik, A.J.; Friedel, M.G.; Carell, T.
Characterization of a new thermophilic spore photoproduct lyase from Geobacillus stearothermophilus (SplG) with defined lesion containing DNA substrates (2006), J. Biol. Chem., 281, 36317-36326.

Search for more literature for 4.1.99.14

Cloned(Commentary)

Cloned (Comment)	Organism
functional expression of N-terminally His6-tagged enzyme in Escherichia coli strain pLysS (DE3)	Geobacillus stearothermophilus

Inhibitors

Inhibitors	Comment	Organism	Structure
aza-S-adenosyl-L-methionine	complete inhibition	Geobacillus stearothermophilus

Metals/Ions

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/ Products (Substrates)

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

Organism	UniProt	Comment	Textmining
Geobacillus stearothermophilus	-	gene splG	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His6-tagged enzyme from Escherichia coli strain pLysS (DE3) by nickel affinity chromatography and gel filtration	Geobacillus stearothermophilus

Renatured (Commentary)

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

Source Tissue	Comment	Organism	Textmining

Specific Activity [micromol/min/mg]

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 and Products (Substrate)

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

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

Synonyms	Comment	Organism
More	the thermophilic spore photoproduct lyase belongs to the family of radical S-adenosylmethionine enzymes	Geobacillus stearothermophilus
SplG	-	Geobacillus stearothermophilus

Cofactor

Cofactor	Comment	Organism	Structure
S-adenosyl-L-methionine	-	Geobacillus stearothermophilus

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Release 2023.1 (January 2023)