Figure 6: Enzymatic activity of nmt1a gene product. (A) The peptide substrate (GARASVLSK-biotin), myristoyl-CoA and bacterial cell lysate containing NUS-zNMT1 or NUS tag were mixed and reacted at 30 ºC for 1 hour. After collection of the reacted peptide, MS of the peptide were acquired. (B) The recombinant NUS-zNMT1 protein in E.coli lysate was purified by His tag affinity chromatography. The purified sample was reacted with HRV3C protease to removal of NUS tag. Purification of zNMT1 at each process was confirmed by SDS-PAGE followed by CBB staining. (C) The purified recombinant zNMT1 protein was reacted with substrate peptide labeled with biotin as indicated, after linkage between zNMT1 and its substrate peptide. zNMT1-substrate complex in each samples were analyzed by SDS-PAGE followed by immunoblotting with anti-biotin antibody.