Wright, MH, Paape, D, Price, HP, Smith, DF and Tate, EW (2016) Global profiling and inhibition of protein lipidation in vector and host stages of the sleeping sickness parasite Trypanosoma brucei. ACS Infectious Diseases, 2 (6). pp. 427-441. ISSN 2373-8227

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Abstract

The enzyme N-myristoyltransferase (NMT) catalyzes the essential fatty acylation of substrate proteins with myristic acid in eukaryotes and is a validated drug target in the parasite Trypanosoma brucei, the causative agent of African trypanosomiasis (sleeping sickness). N-Myristoylation typically mediates membrane localization of proteins and is essential to the function of many. However, only a handful of proteins are experimentally validated as N-myristoylated in T. brucei. Here, we perform metabolic labeling with an alkyne-tagged myristic acid analogue, enabling the capture of lipidated proteins in insect and host life stages of T. brucei. We further compare this with a longer chain palmitate analogue to explore the chain length-specific incorporation of fatty acids into proteins. Finally, we combine the alkynyl-myristate analogue with NMT inhibitors and quantitative chemical proteomics to globally define N-myristoylated proteins in the clinically relevant bloodstream form parasites. This analysis reveals five ARF family small GTPases, calpain-like proteins, phosphatases, and many uncharacterized proteins as substrates of NMT in the parasite, providing a global view of the scope of this important protein modification and further evidence for the crucial and pleiotropic role of NMT in the cell.

Item Type: Article
Additional Information: This is the accepted author manuscript (AAM). The final published version (version of record) is available online via ACS at http://dx.doi.org/10.1021/acsinfecdis.6b00034 Please refer to any applicable terms of use of the publisher.
Uncontrolled Keywords: human African trypanosomiasis, N-myristoylation, chemical proteomics, click chemistry, protein lipidation, target validation
Subjects: Q Science > QD Chemistry
Divisions: Faculty of Natural Sciences > School of Life Sciences
Depositing User: Symplectic
Date Deposited: 05 May 2016 14:28
Last Modified: 18 Jun 2018 11:31
URI: http://eprints.keele.ac.uk/id/eprint/1706

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