N-Lauroylation during the Expression of Recombinant N-Myristoylated Proteins: Implications and Solutions.

ABSTRACT: Incorporation of myristic acid onto the N terminus of a protein is a crucial modification that promotes membrane binding and correct localization of important components of signaling pathways. Recombinant expression of N-myristoylated proteins in Escherichia coli can be achieved by co-expressing yeast N-myristoyltransferase and supplementing the growth medium with myristic acid. However, undesired incorporation of the 12-carbon fatty acid lauric acid can also occur (leading to heterogeneous samples), especially when the available carbon sources are scarce, as it is the case in minimal medium for the expression of isotopically enriched samples. By applying this method to the brain acid soluble protein 1 and the 1-185 N-terminal region of c-Src, we show the significant, and protein-specific,

SUBMITTER: Flamm AG 

PROVIDER: S-EPMC4736449 | biostudies-literature | 2016 Jan

REPOSITORIES: biostudies-literature