Thinon, E. et al. Nat. Commun. 5, 4919 (2014).

Little is known about the extent of N-myristoylation in the human proteome. This co- and post-translational modification, which has been implicated in diseases ranging from bacterial infection to epilepsy to cancer, is catalyzed by the N-myristoyltransferase enzyme that transfers myristate to the N-terminal glycine residue of a substrate protein. Thinon et al. now report a chemical proteomic approach that is highly specific for detecting this modification. They used an alkyne-tagged myristate analog that is metabolically incorporated by cells, enabling N-myristoylated proteins to be enriched via affinity purification for mass spectrometry analysis. By treating one population of myristate analog–labeled human cells with increasing concentrations of an N-myristoyltransferase inhibitor and performing a quantitative proteomic analysis, they could determine which putative N-myristoylated proteins were true N-myristoyltransferase substrates.