DNA Repair Enzymes

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  • NEB carries an array of reliable DNA repair enzymes for use in multiple applications.

    Properties of DNA Repair Enzymes

    Enzyme Substrate Cleavage site Termini created from cleavage
    5' terminus 3' terminus
    APE 1 AP sites 1st phosphodiester bond 5´ to the lesion dR5P OH
    Endo III Oxidized Pyrimidines, AP sites 1st phosphodiester bond 3´ to the lesion P PA
    Endo IV AP sites 1st phosphodiester bond 5´ to the lesion dR5P OH
    Endo V Deoxyinosine, mismatches, hairpin/unpaired loop, flaps, pseudo Y structures 2nd or 3rd phosphodiester bond 3´ to the lesion P OH
    Endo VIII Oxidized Pyrimidines, AP sites 1st phosphodiester bond both 5´ and 3´ to the lesion P P
    Fpg Oxidized Purines, AP sites 1st phosphodiester bond both 5´ and 3´ to the lesion P P
    hOGG1 Oxidized Purines, AP sites 1st phosphodiester bond 3´ to the lesion P PA
    hNEIL1 Oxidized Pyrimidines and Purines, AP sites 1st phosphodiester bond both 5´ and 3´ to the lesion P P
    T7 Endo I Cruciform, Holliday junctions, mismatches, herteroduplexes 1st, 2nd or 3rd phosphodiester bond 5´ to the mismatches P OH
    T4 PDG Pyrimidine Dimers N-glycosidic bond of the 5´ Thymine of the dimer and the 1st phosphodiester bond 3´ to the AP site P* PA
    UDG & Afu UDG Deoxyuridine N-glycosidic bond AP site (no break)
    hSMUG1 Deoxyuridine, 5-hyroxyuracil, 5-hydroxymethyluracil, 5-formyluracil N-glycosidic bond AP site (no break)
    hAAG Deoxyinosine, Alkylated purines N-glycosidic bond AP site (no break)

    *A pyrimidine dimer still covalently attached.

    Table Legend:
    AP apurinic/apyrimidinic sites
    P phosphate
    OH hydroxyl
    dR5P deoxyribose-5´-phosphate
    PA 3´-phospho-α, β-unsaturated aldehyde

    DNA Repair Glycosylases on Various Damaged Bases

      Double-stranded DNA Oligos (34-mers)
    Repair
    Enzymes
    ENZYME AP:A DHT:A 5-hmU:A 5-hmU:G I:T 6-MeA:T 8-0G:C 8-0G:G U:A U:G THYMINE GLYCOL:A
    APE 1 ++++ + - - - - - - - - -
    Endo III ++++ + - - - - - - - - +
    Tma Endo III ++++ ++ - - - - + + - - ++
    Endo IV ++++ + - - - - - - - - -
    Tth Endo IV ++++ + - - - - - + - - -
    Endo V* +++ + + + ++++ + ++ + + ++++ ++
    Endo VIII ++++ ++ - - - - - - - - +++
    Fpg + + - - - - ++++ ++++ - - +
    hAAG - - - - ++++ - - - - - -
    hNEIL1 ++++ ++ - - - - + + - - ++
    hOGG1 ++ - - - - - ++++ + - - -
    T4 PDG ++++ - - - - - - - - - -
    UDG N/A - - - - - - - ++++ + -
    Afu UDG N/A - - - - - - - ++++ + -
    hSMUG1 N/A - +++ +++ - - - - ++++ ++++ -

    Standard reaction conditions were used to titer the enzymes with the alternate base
    *Nicks only, does not remove damage

    Single-stranded DNA Oligos (34-mers)
    Repair
    Enzymes
    ENZYME AP DHT 5-hmU I 6-MeA 8-0G U THYMINE GLYCOL:A
    APE 1 ++ - - - - - - -
    Endo III ++ - - - - - - -
    Tma Endo III ++ + - - - - - -
    Endo IV - - - - - - - -
    Tth Endo IV - - - - - - - -
    Endo V + - - ++++ - + - -
    Endo VIII +++ - - - - - - -
    Fpg + + - - - + - +
    hAAG - - - + - - - -
    hNEIL1 + + - - - - - +
    hOGG1 ++ - - - - + - -
    T4 PDG - - - - - - - -
    UDG N/A - - - - - ++++ -
    Afu UDG N/A - - - - - ++++ -
    hSMUG1 N/A - ++ - - - +++ -

    Table Legend:
    AP apurinic/apyrimidinic site. The AP site is created by treating a uracil containing oligo with UDG.
    DHT 5,6-dihydrothymine 
    5-hmU 5-hydroxymethyluracil
    I Inosine 
    6-MeA 6-methyladenine 
    8-OG 8-oxoguanine 
    U uridine
    AP:A apurinic/apyrimidinic site base paired with adenine
    DHT:A 5,6 dihydrothymine base paired with an adenine
    5-hmU:A 5-hydroxymethyluracil base paired with an adenine
    5-hmU:G 5-hydroxymethyluracil base paired with a guanine
    I:T Inosine base paired with a thymine
    6-MeA:T 6-methyladenine base paired with a thymine
    8-OG:C 8-oxoguanine base paired with a cytosine
    8-OG:G 8-oxoguanine base paired with a guanine
    U:A uridine base paired with an adenine
    U:G uridine base paired with a guanine

    Level of Repair:
    ++++100%
    +++ 50%
    ++ 10% –25%
    + <10%
    – no detectable enzyme activity (<0.7%) 
    N/A not applicable