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Biochemical and structural analysis of N-myristoyltransferase mediated protein tagging

Methods Enzymol .. 2024-03; 
Paul Monassa , Frédéric Rivière , Cyril Dian , Frédéric Frottin , Carmela Giglione , Thierry Meinnel
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Peptide Synthesis Recombinant TEV (rTEV, EC 3.4. 22.44) protease (Genscript, Z03030,10U/μL); rTEV contains the 231 amino acid sequence of TEV fused toan N-terminal His tag. Custom octapeptides of any sequence are all from GenScript Biotech (Rijswijk, Netherlands). Get A Quote

摘要

N-terminal myristoylation is an essential eukaryotic modification crucial for cellular homeostasis in the context of many physiological processes. Myristoylation is a lipid modification resulting in a C14 saturated fatty acid addition. This modification is challenging to capture due to its hydrophobicity, low abundance of target substrates, and the recent discovery of unexpected NMT reactivity including myristoylation of lysine side chains and N-acetylation in addition to classical N-terminal Gly-myristoylation. This chapter details the high-end approaches developed to characterize the different features of N-myristoylation and its targets through in vitro and in vivo labeling.

关键词

Acetylation; Fatty acid; Glycine; Lysine; Myristoylation; N-myristoyltransferase; N-terminus; Protein acylation; Protein modification.