Active N-Myristoyltransferase 1 (NMT1)

N-Myristoyltransferase 1 (NMT1) is an indispensable eukaryotic enzyme that belongs to the NMT family. NMT1 can catalyses the transfer of myristoyl groups to the amino acid terminal residues of numerous proteins. This catalytic process is required for the growth and development of many eukaryotes and viruses. Furthermore, it is recognized that N-myristoylation can also occur posttranslationally on an internal glycine exposed by caspase cleavage during apoptosis. Thus a functional binding ELISA assay was conducted to detect the interaction of recombinant human NMT1 and recombinant human CASP3. Briefly, biotin-linked NMT1 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 ul were then transferred to CASP3-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST 3 times and incubation with Streptavidin-HRP for 30min, then wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50 µl stop solution to the wells and read at 450 nm immediately. The binding activity of recombinant human NMT1 and recombinant human CASP3 was shown in Figure 1, the EC50 for this effect is 0.01 ug/mL.