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

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

doi:10.1016/j.chembiol.2015.01.003

Megan H Wright, Daniel Paape + Show 4 more

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https://doi.org/10.1016/j.chembiol.2015.01.003

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Journal: Chemistry & Biology	Publication Date: Feb 26, 2015
Citations: 128	License type: cc-by

Affiliation: Imperial College London, University of York

Abstract

SummaryN-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.

Highlights

Over 300 million people are at risk from the leishmaniases, a spectrum of neglected tropical parasitic diseases strongly associated with poverty, which cause around 30,000 deaths annually (Alvar et al, 2012)
A small number of NMT substrates have been characterized in Leishmania, and a global picture of N-myristoylation is lacking
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

Summary

Introduction

Over 300 million people are at risk from the leishmaniases, a spectrum of neglected tropical parasitic diseases strongly associated with poverty, which cause around 30,000 deaths annually (Alvar et al, 2012). The parasite is transmitted by sand fly vectors, which inoculate the promastigote forms into the human host. They are taken up by phagocytic cells, mainly macrophages, and transform into the obligate intracellular amastigotes, which replicate and can infect other phagocytes. There are currently few drugs to treat the leishmaniases, and those that are available have problems of toxicity, teratogenicity, high cost, difficult administration, and/or parasite resistance. These drugs work via nonspecific antibiotic effects, such as disruption of cell membranes, or their mode of action is unknown (Tate et al, 2014). Novel drugs with a defined mechanism of action, high efficacy and low toxicity are urgently required to combat these devastating diseases

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