Post-translational myristoylation during cell death

Dale David Orr Martin
Myristoylation involves the addition of a 14-carbon fatty acid to the N-terminal glycine of proteins by N-myristoyltransferase. Myristoylation promotes protein-membrane and protein-protein interactions that are crucial for the function of myristoylated proteins. Myristoylation occurs cotranslationally on the nascent polypeptide following the removal of the initiator methionine or post-translationally following the proteolytic exposure of an N-terminal glycine. Herein, we describe how we
more » ... bio-orthogonal myristate analogs that are incorporated into proteins at N-terminal glycines in an Nmyristoyltransferase dependent manner, and subsequently, are chemoselectively ligated to various affinity tags that allowed the facile detection of the myristoylated proteins. These methods allowed the detection of myristoylated proteins by western blotting with exposure times ranging from seconds to minutes (~1-5 million faster compared to the incorporation of radioactive myristate into proteins). Ultimately, we identified 7 post-translationally myristoylated proteins during apoptosis. These include the following caspase cleaved protein products: cell division control protein 6 homolog, cytoplasmic dyneinintermediate chain 2A, Huntingtin (Htt), microtubule-actin crosslinking factor 1, the apoptotic regulator induced myeloid leukemia cell
doi:10.7939/r36x2s fatcat:4xkbqrpozbcl7gpywa54v5qxwm