The majority of eukaryotic proteins undergo amino-terminal (N-terminal) acetylation. This plays an important role in stabilising the proteins, preventing cleavage by proteases and modifying protein function.
E. coli usually lack the biological machinery required for N-terminal acetylation of proteins. In some cases this can cause dramatic differences in the structure and function of recombinant proteins produced in bacterial systems. However, the necessary machinery for N-terminal acetylation (mostly NatA and NatB) can itself be produced recombinantly in bacteria.
This led us to develop plasmids which include the DNA for these acetylation complexes. These can simply be co-transformed into bacteria alongside your own plasmids. The bacteria will produce the acetylation complex and then both express and acetylate your protein of interest.
All of our published plasmids are available to academics and nonprofits on addgene. For commercial uses please get in touch.
Our plasmids
We have developed two plasmids for N-terminal acetylation:
N-terminal acetylation in practice
We have used this system in the past to produce a variety of proteins which require N-terminal acetylation for correct folding and function. These include tropomyosin, synuclein and calmodulin (read the paper in FEBS letters).