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Literature summary for 3.2.2.23 extracted from

  • Rabow, L.E.; Kow, Y.W.
    Mechanism of action of base release by Escherichia coli Fpg protein: Role of lysine 155 in catalysis (1997), Biochemistry, 36, 5084-5096.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression of K155A mutant and wild-type Fpg in Escherichia coli BL21(DE3) and BLR(DE3) Escherichia coli

Protein Variants

Protein Variants Comment Organism
K155A mutant with 50fold decreased activity with 7-hydro-8-oxoguanine-DNA as substrate, only 3-4fold decreased activity with 7-methylformamidopyrimidine-DNA, increased AP lyase activity Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0000009
-
DNA pH 7.5, 20°C, 24-oligomer DNA containing apurinic/apyrimidinic sites, lyase activity, K155A mutant Fpg Escherichia coli
0.0000069
-
DNA containing 7-hydro-8-oxoguanine residues pH 7.5, 37°C, 24-oligomer, wild-type Fpg Escherichia coli
0.0000075
-
DNA containing 7-hydro-8-oxoguanine residues pH 7.5, 37°C, 24-oligomer, K155A mutant Fpg Escherichia coli
0.000019
-
DNA pH 7.5, 20°C, 24-oligomer DNA containing AP sites, lyase activity, wild-type Fpg Escherichia coli
0.000023
-
DNA containing ring-opened N7-methylguanine residues pH 7.5, 37°C, 23-oligomer, wild-type Fpg Escherichia coli
0.000029
-
DNA containing ring-opened N7-methylguanine residues pH 7.5, 37°C, 23-oligomer, K155A mutant Fpg Escherichia coli

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
35000
-
1 * 35000, SDS-PAGE Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
DNA + H2O Escherichia coli removes oxidized purines from oxidatively damaged DNA ?
-
?
DNA + H2O Escherichia coli DNA base excision repair enzyme ?
-
?

Organism

Organism UniProt Comment Textmining
Escherichia coli P50465 strain 5H11S
-

Purification (Commentary)

Purification (Comment) Organism
wild-type and K155A mutant Fpg, expressed in Escherichia coli BLR(DE3) Escherichia coli

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.054
-
polymeric DNA containing ring-opened N7-methylguanine residues as substrate, wild-type Fpg Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
DNA + H2O with apurinic/apyrimidinic lyase activity, catalyzes beta and delta elimination reactions Escherichia coli ?
-
?
DNA + H2O removes oxidized purines from oxidatively damaged DNA Escherichia coli ?
-
?
DNA + H2O enzyme has N-glycosylase and apurinic/apyrimidinic lyase activity Escherichia coli ?
-
?
DNA + H2O DNA base excision repair enzyme Escherichia coli ?
-
?
DNA containing 2'-deoxy-8-oxonebularine residues + H2O 23mer oligonucleotide containing a single site, very poor substrate Escherichia coli ?
-
?
DNA containing 2,6-diamino-4-hydroxy-5-formamidopyrimidine residues + H2O FapyGua Escherichia coli 2,6-diamino-4-hydroxy-5-formamidopyrimidine + DNA
-
?
DNA containing 4,6-diamino-5-formamidopyrimidine residues + H2O FapyAde Escherichia coli 4,6-diamino-5-formamidopyrimidine + DNA
-
?
DNA containing 7-hydro-8-oxoguanine residues + H2O 24-oligomer, Lys-155 directly interacts with the C8 oxygen of 8-oxopurines involving proton transfer or transient formation of an ion pair between enzyme and substrate, mechanism Escherichia coli DNA + 7-hydro-8-oxoguanine
-
?
DNA containing 7-hydro-8-oxoguanine residues + H2O DNA glycosylase/AP lyase activity Escherichia coli DNA + 7-hydro-8-oxoguanine
-
?
DNA containing ring-opened N7-methylguanine residues + H2O 23-oligomer Escherichia coli 2,6-diamino-4-hydroxy-5-N-methylformamidopyrimidine + DNA
-
?

Subunits

Subunits Comment Organism
monomer 1 * 35000, SDS-PAGE Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.000383
-
DNA containing 7-hydro-8-oxoguanine residues pH 7.5, 37°C, 24-oligomer, K155A mutant Fpg Escherichia coli
0.0012
-
DNA containing ring-opened N7-methylguanine residues pH 7.5, 37°C, 23-oligomer, K155A mutant Fpg Escherichia coli
0.00383
-
DNA containing ring-opened N7-methylguanine residues pH 7.5, 37°C, 23-oligomer, wild-type Fpg Escherichia coli
0.0185
-
DNA containing 7-hydro-8-oxoguanine residues pH 7.5, 37°C, 24-oligomer, wild-type Fpg Escherichia coli
0.0568
-
DNA pH 7.5, 20°C, 24-oligomer DNA containing AP sites, lyase activity, wild-type Fpg Escherichia coli
0.0883
-
DNA pH 7.5, 20°C, 24-oligomer DNA containing AP sites, lyase activity, K155A mutant Fpg Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Escherichia coli