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F3174

Sigma-Aldrich

Fpg Protein from Escherichia coli

≥90% (SDS-PAGE), buffered aqueous glycerol solution, >20,000 units/mg protein, suitable for genomic analysis

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

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About This Item

CAS Number:
Enzyme Commission number:
MDL number:
UNSPSC Code:
12352202
NACRES:
NA.32

biological source

Escherichia coli

Quality Level

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.

application(s)

genomic analysis

shipped in

wet ice

storage temp.

−20°C

Gene Information

Escherichia coli CFT073 ... mutM(1038243)
Escherichia coli K12 ... mutM(946765)

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.

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

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Kim Jantzen et al.
Mutagenesis, 27(6), 693-701 (2012-08-08)
Studies in mono-culture of cells have shown that diesel exhaust particles (DEPs) increase the production of reactive oxygen species (ROS) and oxidative stress-related damage to DNA. However, the level of particle-generated genotoxicity may depend on interplay between different cell types
Lotte Frigaard Mandsberg et al.
FEMS microbiology letters, 324(1), 28-37 (2011-11-19)
Prevention and correction of oxidative DNA lesions in Pseudomonas aeruginosa is ensured by the DNA oxidative repair system (GO). Single inactivation of mutT, mutY and mutM involved in GO led to elevated mutation rates (MRs) that correlated to increased development
S Boiteux et al.
The Journal of biological chemistry, 265(7), 3916-3922 (1990-03-05)
The repair of 2,6-diamino-4-hydroxy-5-N-methyl-formamidopyrimidine (Fapy) residues in DNA is performed by a Fapy-DNA glycosylase activity which is encoded for by the fpg gene in Escherichia coli. Besides DNA glycosylase activity, this protein, the FPG protein, is endowed with an EDTA-resistant
S Boiteux et al.
The EMBO journal, 6(10), 3177-3183 (1987-10-01)
An Escherichia coli genomic library composed of large DNA fragments (10-15 kb) was constructed using the plasmid pBR322 as vector. From it 700 clones were individually screened for increased excision of the ring-opened form of N7-methylguanine (2-6-diamino-4-hydroxy-5N-methyl-formamidopyrimidine) or Fapy. One
Derrik M Leach et al.
Mutagenesis, 28(5), 507-513 (2013-06-25)
Previously, we have reported the use of a recombinant adenovirus (Ad)-based host cell reactivation (HCR) assay to examine nucleotide excision repair (NER) of UVC-induced DNA lesions in several mammalian cell types. The recombinant non-replicating Ad expresses the Escherichia coli β-galactosidase

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