Application | Comment | Organism |
---|---|---|
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 (Comment) | Organism |
---|---|
cloned in Escherichia coli | Thermus thermophilus |
Crystallization (Comment) | Organism |
---|---|
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 | Comment | Organism |
---|---|---|
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 | UniProt | Comment | Textmining |
---|---|---|---|
Thermus thermophilus | P61497 | - |
- |
Purification (Comment) | Organism |
---|---|
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 | Comment | Organism | Textmining |
---|
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
cyclobutadipyrimidine in DNA | - |
Thermus thermophilus | 2 pyrimidine residues in DNA | - |
? |
Synonyms | Comment | Organism |
---|---|---|
CPD-photolyase | - |
Thermus thermophilus |
DNA photolyase | - |
Thermus thermophilus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Thermus thermophilus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
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 | |
8-iodoflavin | chromophore binding site of Thermus photolyase is reconstited also with a novel synthetic flavin, 8-iodoflavin | Thermus thermophilus | |
flavin | flavin-mononucleotide (FMN), crystal strucutre analysis reveals the binding of flavin mononucleotide as an antenna chromophore | Thermus thermophilus |