ALKBH2 Human

ALKBH2 is produced in Escherichia coli (E. coli) as a single, non-glycosylated polypeptide chain containing 281 amino acids, with a molecular mass of approximately 31.4 kDa . The recombinant protein is fused to a 20 amino acid His-tag at the N-terminus, which facilitates its purification using chromatographic techniques .

The primary function of ALKBH2 is to repair alkylated DNA bases by oxidative dealkylation. This process involves the removal of alkyl groups from damaged DNA bases, thereby restoring the DNA to its undamaged state. ALKBH2 uses molecular oxygen, 2-oxoglutarate, and iron as cofactors to oxidize the alkyl groups, which are subsequently released as aldehydes . This enzyme can process both double-stranded (dsDNA) and single-stranded DNA (ssDNA) substrates, with a strong preference for dsDNA .

ALKBH2 specifically repairs monoalkylated bases such as N1-methyladenine and N3-methylcytosine, as well as higher-order alkyl adducts like 1,N6-ethenoadenine, 3,N4-ethenocytosine, and 1,N2-ethenoguanine . The enzyme locates weakened base pairs, flips the damaged base out of the DNA helix, and accommodates the lesion in its active site for efficient catalysis .

The repair of alkylated DNA is essential for preventing mutations and maintaining genomic integrity. ALKBH2 plays a vital role in protecting cells from the cytotoxic effects of alkylating agents, which can cause DNA damage and lead to various diseases, including cancer . By repairing alkylated DNA, ALKBH2 helps to ensure the proper functioning of cellular processes and the stability of the genome.

Recombinant ALKBH2 is used in various research applications to study DNA repair mechanisms and the effects of alkylating agents on DNA. It is also utilized in drug discovery and development to identify potential therapeutic targets for diseases associated with DNA damage and repair.

The recombinant ALKBH2 protein is typically stored at -20°C for long-term use and at 4°C for short-term use (2-4 weeks). To prevent degradation, it is recommended to add a carrier protein such as 0.1% human serum albumin (HSA) or bovine serum albumin (BSA) and avoid multiple freeze-thaw cycles .