Literature summary extracted from
Klar, T.; Kaiser, G.; Hennecke, U.; Carell, T.; Batschauer, A.; Essen, L.O.
Natural and non-natural antenna chromophores in the DNA photolyase from Thermus thermophilus (2006), Chembiochem, 7, 1798-1806.
Application
EC Number |
Application |
Comment |
Organism |
---|
4.1.99.3 |
biotechnology |
it should be possible to perform wavelength tuning of the Thermus photolyase by using artificial flavin chromophores and photolyase variants whose antenna chromophore-binding sites are reengineered by molecular modeling and mutagenesis |
Thermus thermophilus |
Cloned(Commentary)
EC Number |
Cloned (Comment) |
Organism |
---|
4.1.99.3 |
cloned in Escherichia coli |
Thermus thermophilus |
Crystallization (Commentary)
EC Number |
Crystallization (Comment) |
Organism |
---|
4.1.99.3 |
hexagonal crystals are obtained by the hangingdrop vapor-diffusion method after four days. Coordinates and structure factors of the photolyase-chromophore complexes are deposited in the RCSB protein databank under the accession codes 2J07, 2J08, and 2J09 |
Thermus thermophilus |
Protein Variants
EC Number |
Protein Variants |
Comment |
Organism |
---|
4.1.99.3 |
R46E |
R46E mutant which lacks a conserved arginine in the binding site for the antenna chromophore shows no flavin-mononucleotide and discloses an eightfold lower activity at 450 nm (blue light) wheras at 370 nm (UV-A light) its activity is only three times lower than wildtype enzyme |
Thermus thermophilus |
Organism
EC Number |
Organism |
UniProt |
Comment |
Textmining |
---|
4.1.99.3 |
Thermus thermophilus |
P61497 |
- |
- |
Purification (Commentary)
EC Number |
Purification (Comment) |
Organism |
---|
4.1.99.3 |
cells are disrupted by an emulsifier, followed by heat treatment for 15 min at 65°C and centrifugation. Supernatant is purified by using a Blue Sepharose CL-6B affinity column. Recombinant photolyase elutes at a concentration of 1 M KCl in an increasing salt gradient. Fraction containing the protein are combined and concentrated by ultrafiltration. Gel filtration chromatography with Sephacryl S200-HR as a final step give an overal yield of 35 mg of purified CPD-photolyase per liter cell culture |
Thermus thermophilus |
Source Tissue
EC Number |
Source Tissue |
Comment |
Organism |
Textmining |
---|
Substrates and Products (Substrate)
EC Number |
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
---|
4.1.99.3 |
cyclobutadipyrimidine in DNA |
- |
Thermus thermophilus |
2 pyrimidine residues in DNA |
- |
? |
|
Synonyms
EC Number |
Synonyms |
Comment |
Organism |
---|
4.1.99.3 |
CPD-photolyase |
- |
Thermus thermophilus |
4.1.99.3 |
DNA photolyase |
- |
Thermus thermophilus |
pH Optimum
EC Number |
pH Optimum Minimum |
pH Optimum Maximum |
Comment |
Organism |
---|
4.1.99.3 |
7.5 |
- |
assay at |
Thermus thermophilus |
Cofactor
EC Number |
Cofactor |
Comment |
Organism |
Structure |
---|
4.1.99.3 |
7,8-didemethyl-8-hydroxy-5-deazariboflavin |
chromophore binding site of Thermus photolyase is reconstited also with 7,8-didemethyl-8-hydroxy-5-deazariboflavin (8-HDF). However, in the genome sequence of Thermus thermophilus it is found that the genes essential for the biosynthesis of 8-HDF are missing |
Thermus thermophilus |
|
4.1.99.3 |
8-iodoflavin |
chromophore binding site of Thermus photolyase is reconstited also with a novel synthetic flavin, 8-iodoflavin |
Thermus thermophilus |
|
4.1.99.3 |
flavin |
flavin-mononucleotide (FMN), crystal strucutre analysis reveals the binding of flavin mononucleotide as an antenna chromophore |
Thermus thermophilus |
|