Dioxygenase that repairs alkylated DNA and RNA containing 3-methylcytosine or 1-methyladenine by oxidative demethylation. Has highest activity towards 3-methylcytosine. Has lower activity towards alkylated DNA containing ethenoadenine, and no detectable activity towards 1-methylguanine or 3-methylthymine. Accepts double-stranded and single-stranded substrates. Requires molecular oxygen, alpha-ketoglutarate and iron. Provides extensive resistance to alkylating agents such as MMS and DMS (SN2 agents), but not to MMNG and MNU (SN1 agents). Source: Recombinant protein corresponding to aa1-220 from Caulobacter vibrioides Alpha-ketoglutarate-dependent dioxygenase AlkB homolog fused to 10X His-Tag at N-terminal and Myc-Tag at C-terminal, expressed in E.coli. Molecular Weight: ~31.2kD Amino Acid Sequence: MAVVRRAVAARGLQMIAKPLTVVPGFDVWPGLLDISAQRALVEAVLAGAEQAPFSNYRTAYGKPMSVAMTALGSLGWTSDARGYRYVDRHPETGRPWPDMPPALLDLWTVLGDPETPPDSCLVNLYRDGARMGLHQDRDEADPRFPVLSISLGDTAVFRIGGVNRKDPTRSLRLASGDVCRLLGPARLAFHGVDRILPGSSSLVPGGGRINLTLRRARTA Storage and Stability: May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. Aliquots are stable for 6 months after receipt at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
