Details

Autor(en) / Beteiligte

• Wright, Megan H.
• Paape, Daniel
• Storck, Elisabeth M.
• Serwa, Remigiusz A.
• Smith, Deborah F.
• Tate, Edward W.

Titel

Global Analysis of Protein N-Myristoylation and Exploration of N-Myristoyltransferase as a Drug Target in the Neglected Human Pathogen Leishmania donovani

Ist Teil von

• Chemistry & biology, 2015-03, Vol.22 (3), p.342-354

Ort / Verlag

United States: Elsevier Ltd

Erscheinungsjahr

2015

Link zum Volltext

Quelle

MEDLINE

Beschreibungen/Notizen

• N-Myristoyltransferase (NMT) modulates protein function through the attachment of the lipid myristate to the N terminus of target proteins, and is a promising drug target in eukaryotic parasites such as Leishmania donovani. Only a small number of NMT substrates have been characterized in Leishmania, and a global picture of N-myristoylation is lacking. Here, we use metabolic tagging with an alkyne-functionalized myristic acid mimetic in live parasites followed by downstream click chemistry and analysis to identify lipidated proteins in both the promastigote (extracellular) and amastigote (intracellular) life stages. Quantitative chemical proteomics is used to profile target engagement by NMT inhibitors, and to define the complement of N-myristoylated proteins. Our results provide new insight into the multiple pathways modulated by NMT and the pleiotropic effects of NMT inhibition. This work constitutes the first global experimental analysis of protein lipidation in Leishmania, and reveals the extent of NMT-related biology yet to be explored for this neglected human pathogen. [Display omitted] •Alkyne-tagged probes for protein lipidation are applied in Leishmania donovani•A global profile of protein lipidation in two life stages is presented•N-Myristoylated proteome defined by chemical knockdown and quantitative proteomics•Evidence for NMT as a drug target in the leishmaniases Wright et al. use metabolic tagging with an alkyne-myristate analog, click chemistry, and proteomics to identify lipidated proteins in two life stages of the parasite Leishmania donovani. Quantitative profiling of N-myristoyltransferase inhibition and identification of lipidation sites define the N-myristoylated proteome in this human pathogen.