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Literature summary extracted from

  • Munshi, S.; Rajamoorthi, A.; Stanley, R.J.
    Characterization of a cold-adapted DNA photolyase from C. psychrerythraea 34H (2017), Extremophiles, 21, 919-932 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.1.99.3 recombinant expression of CpPL in Escherichia coli BL21(DE3) in inclusion bodies, as His6-tagged protein in Escherichia coli Arctic Express (DE3) cells, and as MBP-fusion protein in Escherichia coli strain BL21. Recombinant CpPL (rCpPL) binds two different second cofactor molecules, flavin mononucleotide (FMN) when overexpressed and purified from Escherichia coli BL21 (DE3) inclusion bodies, and a folate (possibly MTHF) when overexpressed and purified from Escherichia coli Arctic Express (DE3) cells as a His6-tagged protein or in strain BL21(DE3) cells as a MBP-fusion protein Colwellia psychrerythraea

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4.1.99.3 cyclobutadipyrimidine (in DNA) Colwellia psychrerythraea
-
2 pyrimidine residues (in DNA)
-
?
4.1.99.3 cyclobutadipyrimidine (in DNA) Colwellia psychrerythraea 34H / ATCC BAA-681
-
2 pyrimidine residues (in DNA)
-
?

Organism

EC Number Organism UniProt Comment Textmining
4.1.99.3 Colwellia psychrerythraea Q485Z2 i.e. Vibrio psychroerythus
-
4.1.99.3 Colwellia psychrerythraea 34H / ATCC BAA-681 Q485Z2 i.e. Vibrio psychroerythus
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.1.99.3 recombinant His6-tagged enzyme CpPL from Escherichia coli strain Arctic Express (DE3) by nickel affinity chromatography, and MBP-fusion CpPL from Escherichia coli strain Arctic Express (DE3) by amylose affinity chromatography Colwellia psychrerythraea

Reaction

EC Number Reaction Comment Organism Reaction ID
4.1.99.3 cyclobutadipyrimidine (in DNA) = 2 pyrimidine residues (in DNA) reaction mechanism Colwellia psychrerythraea

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.1.99.3 cyclobutadipyrimidine (in DNA)
-
Colwellia psychrerythraea 2 pyrimidine residues (in DNA)
-
?
4.1.99.3 cyclobutadipyrimidine (in DNA)
-
Colwellia psychrerythraea 34H / ATCC BAA-681 2 pyrimidine residues (in DNA)
-
?
4.1.99.3 additional information CpPL is fully competent to bind and base flip CPDs, and to repair them when exposed to blue light. rCpPL recognizes and flips out a CPD into its active site, base flipping of the CPD by photolyase is accompanied by a large distortion of the local structure of the DNA duplex around the lesion, including the loss of DNA base stacking. 2-Ap base-flipping assay, overview. Thermodynamically, the apparent lack of rigidity of the chains forming the active site would impart a high degree of conformational entropy to the active site of CpPL Colwellia psychrerythraea ?
-
?
4.1.99.3 additional information CpPL is fully competent to bind and base flip CPDs, and to repair them when exposed to blue light. rCpPL recognizes and flips out a CPD into its active site, base flipping of the CPD by photolyase is accompanied by a large distortion of the local structure of the DNA duplex around the lesion, including the loss of DNA base stacking. 2-Ap base-flipping assay, overview. Thermodynamically, the apparent lack of rigidity of the chains forming the active site would impart a high degree of conformational entropy to the active site of CpPL Colwellia psychrerythraea 34H / ATCC BAA-681 ?
-
?

Subunits

EC Number Subunits Comment Organism
4.1.99.3 More predicted three-dimensional structure of CpPL Colwellia psychrerythraea

Synonyms

EC Number Synonyms Comment Organism
4.1.99.3 cold-adapted DNA photolyase
-
Colwellia psychrerythraea
4.1.99.3 CpPL
-
Colwellia psychrerythraea
4.1.99.3 DNA photolyase
-
Colwellia psychrerythraea
4.1.99.3 PhrB
-
Colwellia psychrerythraea

Cofactor

EC Number Cofactor Comment Organism Structure
4.1.99.3 FAD involved in catalysis, cold-adapted DNA photolyase binds a catalytic flavin adenine dinucleotide (FAD) cofactor noncovalently. UV/Vis and fluorescence spectroscopy reveal that the FAD-binding site in this psychrophilic protein is unique compared to meso/thermophilic PLs. FAD-binding pocket of the CpPL model, overview Colwellia psychrerythraea
4.1.99.3 methenyltetrahydrofolate MTHF, the molecule is bound as an antenna molecule and found in substoichiometric amounts Colwellia psychrerythraea
4.1.99.3 additional information recombinant CpPL (rCpPL) binds two different second cofactor molecules, flavin mononucleotide (FMN) when overexpressed and purified from Escherichia coli BL21(DE3) inclusion bodies, and a folate (possibly MTHF) when overexpressed and purified from Escherichia coli Arctic Express (DE3) cells as a His6-tagged protein or in strain BL21-DE3 cells as a maltose-binding-protein fusion protein. CpPL might be somewhat promiscuous in antenna cofactor binding Colwellia psychrerythraea

General Information

EC Number General Information Comment Organism
4.1.99.3 additional information homology analysis of PL protein structures spanning 70°C in growth temperature supports the data that the structure of cold-adapted DNA photolyase CpPL is quite different from warm-adapted DNA photolyases. Homology modeling of CpPL using CPD-PL from Sulfolobus tokodaii (StPL, 2E0I. PDB, chain A) as a template Colwellia psychrerythraea