Global analysis of protein N-myristoylation and exploration of N-myristoyltransferase as a drug target in the neglected human pathogen Leishmania donovani.
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.
Department of Chemistry, Imperial College London, London SW7 2AZ, UK. Electronic address: megan.wright@tum.de.
Megan H
MH
Wright
Centre for Immunology and Infection, Department of Biology, University of York, York YO10 5DD, UK.
Daniel
D
Paape
Department of Chemistry, Imperial College London, London SW7 2AZ, UK.
Elisabeth M
EM
Storck
Department of Chemistry, Imperial College London, London SW7 2AZ, UK.
Remigiusz A
RA
Serwa
Centre for Immunology and Infection, Department of Biology, University of York, York YO10 5DD, UK.
Deborah F
DF
Smith
Department of Chemistry, Imperial College London, London SW7 2AZ, UK. Electronic address: e.tate@imperial.ac.uk.
Edward W
EW
Tate
D000217:Acyltransferases / Q000032:analysis / Q000378:metabolism*; D000818:Animals; D006801:Humans; D007893:Leishmania donovani / Q000737:chemistry / Q000378:metabolism*; D007896:Leishmaniasis / Q000469:parasitology; D051379:Mice; D008807:Mice, Inbred BALB C; D018345:Mice, Knockout; D009226:Myristates / Q000032:analysis / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D040901:Proteomics / Q000379:methods; D015800:Protozoan Proteins / Q000032:analysis / Q000378:metabolism*
D009226:Myristates; D015800:Protozoan Proteins; D000217:Acyltransferases; C050480:glycylpeptide N-tetradecanoyltransferase