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Literature summary for 4.1.99.3 extracted from

  • Gindt, Y.M.; Edani, B.H.; Olejnikova, A.; Roberts, A.N.; Munshi, S.; Stanley, R.J.
    The missing electrostatic interactions between DNA substrate and Sulfolobus solfataricus DNA photolyase what is the role of charged amino acids in thermophilic DNA binding proteins? (2016), J. Phys. Chem. B, 120, 10234-10242 .
    View publication on PubMed
Search for more literature for 4.1.99.3

Cloned(Commentary)

Cloned (Comment) Organism
gene phrB, recombinant expression in Escherichia coli strain Rosetta 2 (DE3) Saccharolobus solfataricus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information thermodynamic and kinetic study, overview Saccharolobus solfataricus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
cyclobutadipyrimidine (in DNA) Saccharolobus solfataricus
-
 2 pyrimidine residues (in DNA)
-
 ?
cyclobutadipyrimidine (in DNA) Saccharolobus solfataricus DSM ATCC 35092 / DSM 1617 / JCM 11322 / P2
-
 2 pyrimidine residues (in DNA)
-
 ?

Organism

Organism UniProt Comment Textmining
Saccharolobus solfataricus F9VNB1 i.e. Sulfolobus solfataricus
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Saccharolobus solfataricus DSM ATCC 35092 / DSM 1617 / JCM 11322 / P2 F9VNB1 i.e. Sulfolobus solfataricus
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme from Escherichia coli strain Rosetta 2 (DE3) by desalting gel filtration, Cibacron Blue 3GA affinity and heparin affinity chromatography, and ultrafiltration Saccharolobus solfataricus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
cyclobutadipyrimidine (in DNA)
-
 Saccharolobus solfataricus 2 pyrimidine residues (in DNA)
-
 ?
cyclobutadipyrimidine (in DNA) the substrate used in binding experiments, UV-p(dT)10 (denoted as ssDNA), is a single strand oligothymidylate with an average of a single CPD lesion randomly arranged on the 10mer Saccharolobus solfataricus 2 pyrimidine residues (in DNA)
-
 ?
cyclobutadipyrimidine (in DNA)
-
 Saccharolobus solfataricus DSM ATCC 35092 / DSM 1617 / JCM 11322 / P2 2 pyrimidine residues (in DNA)
-
 ?
cyclobutadipyrimidine (in DNA) the substrate used in binding experiments, UV-p(dT)10 (denoted as ssDNA), is a single strand oligothymidylate with an average of a single CPD lesion randomly arranged on the 10mer Saccharolobus solfataricus DSM ATCC 35092 / DSM 1617 / JCM 11322 / P2 2 pyrimidine residues (in DNA)
-
 ?
additional information the first step in the repair mechanism: substrate recognition and binding is s measured by isothermal titration calorimetry Saccharolobus solfataricus ?
-
 ?
additional information the first step in the repair mechanism: substrate recognition and binding is s measured by isothermal titration calorimetry Saccharolobus solfataricus DSM ATCC 35092 / DSM 1617 / JCM 11322 / P2 ?
-
 ?

Synonyms

Synonyms Comment Organism
DNA photolyase
-
 Saccharolobus solfataricus
PhrB
-
 Saccharolobus solfataricus
SsPL
-
 Saccharolobus solfataricus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
45
-
 recombinant enzyme Saccharolobus solfataricus

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
25 50 the activity increases slightly from 25-45°C. At 50°C under the solvent conditions of the assay, the SsPL appears to denature after repair of the DNA substrate is mostly complete Saccharolobus solfataricus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
 assay at Saccharolobus solfataricus

Cofactor

Cofactor Comment Organism Structure
FAD enzyme SsPL is an unusual photolyase in that it contains two FAD cofactors. One FAD cofactor is part of the active site of the protein and required for both DNA binding and repair. The second cofactor, the putative accessory chromophore, may play a role as a light-harvesting pigment, it is always present in the fully oxidized FAD state. The active site cycles between FADH-, the fully reduced form required for activity, and FADH radical, the one-electron oxidized or semiquinone form, SsPL is isolated with the active site mainly in the FADH· state. The accessory FAD does not appear to readily undergo any reduction-oxidation chemistry, and it is always found in the fully oxidized state Saccharolobus solfataricus

General Information

General Information Comment Organism
additional information the enzymatic activity and thermodynamics of substrate binding for the enzyme from Sulfolobus solfataricus are directly compared to the enzyme from Escherichia coli, overview. Turnover numbers and catalytic activity are virtually identical, but organic co-solvents may be necessary to maintain activity of the thermophilic protein at higher temperatures. UV-damaged DNA binding to the thermophilic protein is less favorable by about 2 kJ/mol. The enthalpy of binding is about 10 kJ/mol less exothermic for the thermophile, but the amount and type of surface area buried upon DNA binding appears to be somewhat similar. The most important finding is observed when ionic strength studies are used to separate binding interactions into electrostatic and nonelectrostatic contributions, DNA binding to the thermophilic protein appears to lack the electrostatic contributions observed with the mesophilic protein. Reported differences between mesophilic and thermophilic enzymes include an increase in the number of ion pairs/salt bridges, better packing of hydrophobic amino acids, and increased hydrogen bonding for the thermophilic proteins. Comparison of the enthalpy of binding. Analysis of enzyme-substrate interactions, overview Saccharolobus solfataricus
physiological function DNA photolyase is a structure-specific DNA repair enzyme that reverses one of the most common types of UV damage in DNA molecules, the cis-syn cyclobutylpyrimidine dimer (CPD) Saccharolobus solfataricus