In order to determine the amino-terminal sequence requirements for protein N-myristoylation, site-directed mutagenesis of the N-terminal region was performed using tumor necrosis factor (TNF) mutants as model substrate proteins. Subsequently, the susceptibility of thesemutants to protein N-myristoylation was evaluated by metabolic labe-ling in anin vitrotranslation systemusing rabbit reticulocyte lysate. A TNF mutant having the sequence MGAAAAA AAA at its N-terminus was used as the starting sequence to identify elements critical for protein N-myristoylation
