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Literature summary for 3.2.2.17 extracted from

  • McCullough, A.K.; Romberg, M.T.; Nyaga, S.; Wie, Y.; Wood, T.G.; Taylor, J.S.; Van Etten, J.L.; Dodson, M.L.; Lloyd, R.S.
    Characterization of a novel cis-syn and trans-symII pyrimidine dimer glycosylase/AP lyase from a eukaryotic algal virus Paramecium bursaria chlorella Virus-1 (1998), J. Biol. Chem., 273, 13136-13142.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
isopropyl-1-thio-beta-D-galactopyranoside induction of the pdg gene Paramecium bursaria Chlorella virus 1

Cloned(Commentary)

Cloned (Comment) Organism
gene cv-pdg encoding T4 endonucleaseV homolog, 41% identical to T4endoV, enhance UV radiation resistance in repair-deficient Escherichia coli BL21, vector pCRII, cloning and overexpression of the gene product, cloned into the Escherichia coli expression plasmid pET11a Paramecium bursaria Chlorella virus 1

Crystallization (Commentary)

Crystallization (Comment) Organism
-
Tequatrovirus T4

General Stability

General Stability Organism
relatively stable and soluble under most conditions Paramecium bursaria Chlorella virus 1

Inhibitors

Inhibitors Comment Organism Structure
NaCl above 40 mM Tequatrovirus T4

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00000014
-
DNA cis-syn 49-bp duplex Paramecium bursaria Chlorella virus 1
0.00000065
-
DNA trans-syn 49-bp duplex Paramecium bursaria Chlorella virus 1
0.00000267
-
DNA cis-syn 12-bp duplex Paramecium bursaria Chlorella virus 1
0.000005
-
DNA wild-type Tequatrovirus T4

Metals/Ions

Metals/Ions Comment Organism Structure
KCl slightly more stimulatory than NaCl Paramecium bursaria Chlorella virus 1
NaCl
-
Micrococcus luteus
NaCl
-
Tequatrovirus T4
NaCl optimal activity at 100-120 mM Paramecium bursaria Chlorella virus 1

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
16000
-
-
Tequatrovirus T4
16000
-
-
Paramecium bursaria Chlorella virus 1
18000
-
-
Tequatrovirus T4
18000
-
uveA, 27% amino acid identity with T4 endonuclease V Micrococcus luteus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Micrococcus luteus only nicking of UV-irradiated DNA, creating phospho diester bond breaks 5' with respect to pyrimidine dimers, protein containing the glycosylase activity also contains the apyrimidinic endonuclease activity, multifunctional enzyme, 2 distinct domains in a single enzyme molecule possess both pyrimidine dimer-DNA glycosylase, and AP endonuclease activity, first the glycosylase cleaves glycosylic bond of one of the pyrimidine residues of a dimer to yield an AP site, then AP endonuclease hydrolyzes the phospho diester bond near the AP site, prefers duplex DNA to single-stranded ?
-
?
additional information Tequatrovirus T4 only nicking of UV-irradiated DNA, creating phospho diester bond breaks 5' with respect to pyrimidine dimers, protein containing the glycosylase activity also contains the apyrimidinic endonuclease activity, multifunctional enzyme, 2 distinct domains in a single enzyme molecule possess both pyrimidine dimer-DNA glycosylase, and AP endonuclease activity, first the glycosylase cleaves glycosylic bond of one of the pyrimidine residues of a dimer to yield an AP site, then AP endonuclease hydrolyzes the phospho diester bond near the AP site, prefers duplex DNA to single-stranded ?
-
?
additional information Paramecium bursaria Chlorella virus 1 only nicking of UV-irradiated DNA, creating phospho diester bond breaks 5' with respect to pyrimidine dimers, protein containing the glycosylase activity also contains the apyrimidinic endonuclease activity, multifunctional enzyme, 2 distinct domains in a single enzyme molecule possess both pyrimidine dimer-DNA glycosylase, and AP endonuclease activity, first the glycosylase cleaves glycosylic bond of one of the pyrimidine residues of a dimer to yield an AP site, then AP endonuclease hydrolyzes the phospho diester bond near the AP site, prefers duplex DNA to single-stranded ?
-
?

Organism

Organism UniProt Comment Textmining
Micrococcus luteus
-
-
-
Paramecium bursaria Chlorella virus 1
-
eukaryotic algal virus, Chlorella virus, PBCV-1
-
Tequatrovirus T4
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Paramecium bursaria Chlorella virus 1
purified from Escherichia coli AB2480 Tequatrovirus T4

Source Tissue

Source Tissue Comment Organism Textmining
infected cell infected Chlorella NC64A Paramecium bursaria Chlorella virus 1
-

Storage Stability

Storage Stability Organism
-80°C stable when stored in 50% glycerol, -20°C stable for several freeze-thaw cycles, 4°C in buffer without glycerol for up to a year without loss of activity Paramecium bursaria Chlorella virus 1

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
DNA + H2O
-
Micrococcus luteus thymine + ?
-
?
DNA + H2O
-
Tequatrovirus T4 thymine + ?
-
?
DNA + H2O cis-syn pyrimidine dimer specific Paramecium bursaria Chlorella virus 1 thymine + ?
-
?
additional information only nicking of UV-irradiated DNA, creating phospho diester bond breaks 5' with respect to pyrimidine dimers, protein containing the glycosylase activity also contains the apyrimidinic endonuclease activity, multifunctional enzyme, 2 distinct domains in a single enzyme molecule possess both pyrimidine dimer-DNA glycosylase, and AP endonuclease activity, first the glycosylase cleaves glycosylic bond of one of the pyrimidine residues of a dimer to yield an AP site, then AP endonuclease hydrolyzes the phospho diester bond near the AP site, prefers duplex DNA to single-stranded Micrococcus luteus ?
-
?
additional information only nicking of UV-irradiated DNA, creating phospho diester bond breaks 5' with respect to pyrimidine dimers, protein containing the glycosylase activity also contains the apyrimidinic endonuclease activity, multifunctional enzyme, 2 distinct domains in a single enzyme molecule possess both pyrimidine dimer-DNA glycosylase, and AP endonuclease activity, first the glycosylase cleaves glycosylic bond of one of the pyrimidine residues of a dimer to yield an AP site, then AP endonuclease hydrolyzes the phospho diester bond near the AP site, prefers duplex DNA to single-stranded Tequatrovirus T4 ?
-
?
additional information only nicking of UV-irradiated DNA, creating phospho diester bond breaks 5' with respect to pyrimidine dimers, protein containing the glycosylase activity also contains the apyrimidinic endonuclease activity, multifunctional enzyme, 2 distinct domains in a single enzyme molecule possess both pyrimidine dimer-DNA glycosylase, and AP endonuclease activity, first the glycosylase cleaves glycosylic bond of one of the pyrimidine residues of a dimer to yield an AP site, then AP endonuclease hydrolyzes the phospho diester bond near the AP site, prefers duplex DNA to single-stranded Paramecium bursaria Chlorella virus 1 ?
-
?

Synonyms

Synonyms Comment Organism
Chlorella virus pyrimidine dimer glycosylase cv-PDG Paramecium bursaria Chlorella virus 1
pyrimidine dimer-specific glycosylase/AP lyase
-
Paramecium bursaria Chlorella virus 1

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
90
-
thermostable, difficult to completely heat inactivate, retains 10% activity after 20 min at 90°C Paramecium bursaria Chlorella virus 1

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6 8 pH optimum for the 2 enzymatic activities differs, AP lyase function at pH 6.5, becoming inactive at pH 8 Tequatrovirus T4
6 8 pH optimum for the 2 enzymatic activities differs, AP lyase function at pH 6.5, becoming inactive at pH 8 Paramecium bursaria Chlorella virus 1

pH Range

pH Minimum pH Maximum Comment Organism
6 8
-
Paramecium bursaria Chlorella virus 1