Abstract: Human Alkbh1 is a member of the AlkB family of proteins. Like other AlkB family proteins, hAlkbh1 is a Fe²⁺ and alpha-ketoglutarate-dependent dioxygenase that catalyzes oxidative demethylation of modified nucleotides and amino acids. hAlkbh1 shows activity against six different substrates such as the m⁶A in ssDNA/ bulge or bubble DNA, m³C in DNA/RNA, methylated lysine in histone H2A, m¹A in mitochondrial tRNA for the demethylation, oxidation of m⁵C in mitochondrial tRNA and lyase cleavage of AP sites in DNA. Also, hAlkbh1 has significant involvement in many cancers, metastasis, developmental defects, hypothermia, etc. Since it is established that protein conformational dynamics play an important role in regulation of AlkB enzymes, my PhD work focuses on the characterization of hAlkbh1 functional dynamics. In this work-in-progress seminar I will present my progresses in (i) the optimization of a protein expression and purification protocol for hAlkbh1, (ii) the establishment of protocols to assay the demethylase and AP lyase activities of hAlkbh1, and (iii) the investigation of hAlkbh1 conformational dynamics by solution NMR.