Figure 1: Quantitative chemical proteomic strategy for global profiling of NMT substrates in mammalian cells.
From: Global profiling of co- and post-translationally N-myristoylated proteomes in human cells
(a) N-terminal N-myristoylation occurs co-translationally following action of methionine aminopeptidase (MetAP) at the ribosome, or post-translationally following cleavage by a protease. An N-terminal glycine residue is the only fully conserved element of the substrate. (b) Chemical proteomic workflow. Cells cultured in normal media with YnMyr and NMT inhibitor were lysed and spiked with a fixed quantity of lysate from cells cultured in SILAC media (R10K8 labelled) with YnMyr. Labelled proteins were captured by CuAAC with AzTB, affinity enriched and classified as substrates (or non-substrates) according to the response of enrichment to inhibition determined by quantitative proteomics (nanoLC-MS/MS, MaxQuant, Perseus and GraFit 7.0).
