Mapping the myristome: a tagging-by-substrate approach

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Co- or post-translational long-chain acylation of proteins is known to modulate myriad cellular processes including protein trafficking and the immune response, with the enzymes which regulate these modifications emerging as drug targets for the treatment of cancer and infectious diseases. There is a pressing need to define the complete repertoire of myristoylated proteins, dubbed the ?myristome?, as this is the key prerequisite in understanding the biological roles that this modification contributes to target proteins. We propose to develop a generic method for the identification of all myristoylated proteins by application of Tagging-by-substrate (TbS), a powerful chemistry-based technology that integrates classical metabolic labelling with recently devised ?click? chemistry. The methodology will be applied to map the myristome of human cell lines and in the protozoan parasite species T. brucei, P. falciparum and L. major. The research will generate invaluable data about the as-yet unknown targets of acylation in vivo, present new targets for functional biology studies and greatly enhance the a priori prediction of the myristome from genomic data by elucidating the sequence specificity of N-terminal myristoylation, which is known to extend up to 14 residues from the site of acylation. It will also provide an ideal platform for Dr Tate to build on his existing synthetic chemical skills by acquiring new biological expertise from established leaders in their fields.