F3174

Fpg Protein from Escherichia coli

≥90% (SDS-PAGE), recombinant, expressed in E. coli, buffered aqueous glycerol solution, >20,000 units/mg protein

• Synonym(s): DNA-(apurinic or apyrimidinic site)lyase MutM (APlyase MutM), Fapy-DNAglycosylase, Formamidopyrimidine-DNA glycosylase, Fpg Protein from Escherichia coli, Recombinant, Fapy DNA glycosylase, Formamidopyrimidine DNA glycosylase, MutM
• CAS Number: 78783-53-6
• Enzyme Commission number: 3.2.2.23 (BRENDA, IUBMB)
• MDL number: MFCD02264758
• NACRES: NA.32

Properties

• biological source: Escherichia coli
• Quality Level: 200
• recombinant: expressed in E. coli
• Assay: ≥90% (SDS-PAGE)
• form: buffered aqueous glycerol solution
• specific activity: >20,000 units/mg protein
• mol wt: 30.2 kDa (269 amino acids, predicted from the nucleotide sequence)
• composition: protein, 0.1- 0.3 mg/mL Bradford
• storage condition: (Tightly closed)
• technique(s): nucleic acid detection: suitable
• UniProt accession no.: P05523
• application(s): genomic analysis
• shipped in: wet ice
• storage temp.: −20°C
• Gene Information: Escherichia coli CFT073 ... mutM(1038243) ; Escherichia coli K12 ... mutM(946765)

Description

General description

Formamidopyrimidine-DNA glycosylase (Fpg) is a DNA repair enzyme found in Escherichia coli, which contains one zinc atom. Proximal to its C-terminal, it contains a zinc-finger motif of CC/CC type.

Fpg contains two domains separated by a flexible hinge.

Research area: Cell signaling

Application

Fpg Protein from Escherichia coli has been used for the assessment of DNA oxidative damage using comet assay.

Biochem/physiol Actions

Formamidopyrimidine-DNA glycosylase (Fpg) cleaves double-stranded DNA containing the damaged base 8-oxo-7,8-dihydroguanine. It functions as an N-glycosylase and apurinic/apyrimidinic lyase.

Fpg is a key enzyme in the DNA base excision repair pathway (BER). It catalyses the excision of a broad spectrum of modified purines. Fpg has both DNA glycosylase activity that removes the mutated base and AP-lyase activity that releases ribose leaving both 5′- and 3′-phosphorylated ends in the DNA. The zinc finger motif at its C-terminus is responsible for the DNA binding and AP-lyase activity. In addition, its N-terminal proline acts as a nucleophile to produce a Schiff base intermediate that is essential for enzyme action.

Fpg specifically acts on 3′- and 5′-phosphodiester bonds.

Unit Definition

One unit will cleave 50% of 0.5 pmol of double-stranded DNA oligomer substrate (8-oxoguanine−mutated) in 10 min at 25 °C.

Physical form

Solution in 50% glycerol containing 50 mM potassium HEPES, pH 7.5, 1 mM DTT, 1 mM EDTA, and 200 mM NaCl.

Safety Information

• Storage Class Code: 10 - Combustible liquids
• WGK: WGK 2
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable
• Personal Protective Equipment: dust mask type N95 (US), Eyeshields, Gloves