The E.coli AlkB protein protects against the cytotoxicity of methylating agents by repair of the specific DNA lesions generated in single-stranded DNA. ALKBH2 and ALKBH3 are E. coli AlkB homologs that catalyze the removal of 1-methyladenine and 3-methylcytosine. As dioxygenase, Human ALKBH3 protein mediates demethylation of DNA and RNA containing 1-methyladenosine (m1A). It repairs alkylated DNA containing 1-methyladenosine (m1A) and 3-methylcytosine (m3C) by oxidative demethylation. ALKBH3 has a strong preference for single-stranded DNA. It able to process alkylated m3C within double-stranded regions via its interaction with ASCC3, which promotes DNA unwinding to generate single-stranded substrate needed for ALKBH3. It also acts on RNA. ALKBH3 demethylates N1-methyladenosine (m1A) RNA, an epigenetic internal modification of messenger RNAs (mRNAs) highly enriched within 5'-untranslated regions (UTRs) and in the vicinity of start codons. It requires molecular oxygen, alpha-ketoglutarate and iron.

Full-length human ALKBH3 cDNA (285aa, derived BC0067257) was constructed with codon optimization using gene synthesis technology and expressed with a small T7-His-TEV cleavage site Tag (29aa) fusion at its N-terminal and 11 Poly-Arginine (11R) tag at its C-terminal. It was expressed in E. coli as inclusion bodies. The final product was refolded using our unique “temperature shift inclusion body refolding” technology and chromatographically purified.

Stefansson OA, et al., CpG promoter methylation of the ALKBH3 alkylation repair gene in breast cancer. BMC Cancer 17 (1), 469 (2017)

Kogaki T, et al., TP53 gene status is a critical determinant of phenotypes induced by ALKBH3 knockdown in non-small cell lung cancers
Biochem. Biophys. Res. Commun. 488 (2), 285-290 (2017)

Ueda Y, et al., AlkB homolog 3-mediated tRNA demethylation promotes protein synthesis in cancer cells. Sci Rep 7, 42271 (2017)

1. May be used for ALKBH3 mediated epigenetic pathway regulation study in various cells by intracellular delivery of this ALKBH3-11R protein directly in vitro cell culture
2. May be used for mapping ALKBH3 protein-protein interaction.
3. May be used as specific substrate protein for kinase, and ubiquitin (Sumo pathway) related enzyme functional screening assays.
4. Potential biomarker protein / therapeutic target protein for cancer prognosis and cancer treatment drug development.
5. As native human ALKBH3 immunogen for specific antibody production.

MASMTGGQQMGRGHHHHHHENLYFQGGEFEEKRRRARVQGAWAAPVKSQAIAQPATTAKSHLHQKPGQTWKNKEHHLSDREFVFKEPQQVVRRAPEPRVIEEGVYEISLSPTGVSRVCLYPGFVDVKEADWILEQLCQDVPWKQRTGIREDSILQLTFKKSAPVSGTATAPQSCWYERPSPPHIPGPAILTRTRLWAPESGGGGSPGRRRRRRRRRRR