Cloned (Comment) | Organism |
---|---|
gene splB, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain Tuner(DE3)-pLysS | Clostridium acetobutylicum |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | [4Fe-4S] cluster | Clostridium acetobutylicum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(5R)-5,6-dihydro-5-(thymidin-7-yl)thymidine | Clostridium acetobutylicum | in double-helical DNA | thymidylyl-(3'->5')-thymidylate | - |
? | |
(5R)-5,6-dihydro-5-(thymidin-7-yl)thymidine | Clostridium acetobutylicum ATCC 824 / DSM 792 / JCM 1419 / LMG 5710 / VKM B-1787 | in double-helical DNA | thymidylyl-(3'->5')-thymidylate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Clostridium acetobutylicum | Q97L63 | - |
- |
Clostridium acetobutylicum ATCC 824 / DSM 792 / JCM 1419 / LMG 5710 / VKM B-1787 | Q97L63 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant N-terminally His6-tagged enzyme from Escherichia coli strain Tuner(DE3)-pLysS by nickel affinity chromatography, dialysis under anaerobic conditions, and ultrafiltration | Clostridium acetobutylicum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(5R)-5,6-dihydro-5-(thymidin-7-yl)thymidine | in double-helical DNA | Clostridium acetobutylicum | thymidylyl-(3'->5')-thymidylate | - |
? | |
(5R)-5,6-dihydro-5-(thymidin-7-yl)thymidine | in double-helical DNA | Clostridium acetobutylicum ATCC 824 / DSM 792 / JCM 1419 / LMG 5710 / VKM B-1787 | thymidylyl-(3'->5')-thymidylate | - |
? | |
additional information | solution phase dynamics of the DNA repair enzyme spore photoproduct lyase as probed by H/D exchange with HDX as a probe to explore the solution phase conformational dynamics in SPL upon binding to undamaged DNA and dinucleotide 5R-SPTpT and dinucleoside 5R-SP substrates. A 6-mer oligonucleotide, 5'-GCAAGT-3', is used as substrate, which is cleaved to and complement 5'-ACT and TGC-3', overview. Mass spectrometric substrate and product analyses. Enzyme SPL has low affinity for substrate or DNA in the absence of S-adenosyl-L-methionine | Clostridium acetobutylicum | ? | - |
? | |
additional information | solution phase dynamics of the DNA repair enzyme spore photoproduct lyase as probed by H/D exchange with HDX as a probe to explore the solution phase conformational dynamics in SPL upon binding to undamaged DNA and dinucleotide 5R-SPTpT and dinucleoside 5R-SP substrates. A 6-mer oligonucleotide, 5'-GCAAGT-3', is used as substrate, which is cleaved to and complement 5'-ACT and TGC-3', overview. Mass spectrometric substrate and product analyses. Enzyme SPL has low affinity for substrate or DNA in the absence of S-adenosyl-L-methionine | Clostridium acetobutylicum ATCC 824 / DSM 792 / JCM 1419 / LMG 5710 / VKM B-1787 | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
SPL | - |
Clostridium acetobutylicum |
SplB | - |
Clostridium acetobutylicum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Clostridium acetobutylicum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Clostridium acetobutylicum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
S-adenosyl-L-methionine | SAM, SPL is a radical SAM enzyme, enzyme SPL has low affinity for substrate or DNA in the absence of SAM. SAM causes significant structural perturbations regardless of the integrity of the iron-sulfur cluster, consistent with the observation that SAM binds SPL not only via coordination of the iron-sulfur cluster, but also by interactions such as hydrogen bonding, hydrophobic interactions and salt bridges. The amino group of the methionyl moiety of SAM interacts with Ser142 and Asp143 and the carboxylate moiety interacts with Lys174 and Ser195, while the adenine and ribose groups are held in place via hydrogen bonding interactions with Tyr96, Tyr98, Ala234 and Ala273 | Clostridium acetobutylicum | |
[4Fe-4S] cluster | - |
Clostridium acetobutylicum |
General Information | Comment | Organism |
---|---|---|
additional information | conformational changes associated with cofactor and substrate binding may serve to provide a solvent inaccessible and protected active site to safely catalyze radical reactions | Clostridium acetobutylicum |
physiological function | on exposure of spores to UV radiation, an unique methylene bridged thymine dimer, 5-thyminyl-5,6-dihydrothymine (spore photoproduct or SP) accumulates as the main photoproduct. This accumulated photo-damage is rapidly repaired upon spore germination. Spore photoproduct lyase (SPL) catalyzes the repair of the UV lesion spore photoproduct (SP) in a reaction dependent on S-adenosyl-L-methionine (SAM) | Clostridium acetobutylicum |