Global profiling and inhibition of protein lipidation in vector and host stages of the sleeping sickness parasite Trypanosoma brucei

Wright, M. H., Paape, D. , Price, H. P., Smith, D. F. and Tate, E. W. (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. (doi: 10.1021/acsinfecdis.6b00034) (PMID:27331140) (PMCID:PMC4906374)

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: Articles
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Paape, Dr Daniel
• Authors: Wright, M. H., Paape, D., Price, H. P., Smith, D. F., and Tate, E. W.
• College/School: College of Medical Veterinary and Life Sciences > School of Infection & Immunity
• Journal Name: ACS Infectious Diseases
• Publisher: American Chemical Society
• ISSN: 2373-8227
• ISSN (Online): 2373-8227
• Published Online: 29 April 2016
• Copyright Holders: Copyright © 2016 American Chemical Society
• First Published: First published in ACS Infectious Diseases 2(6):427-441
• Publisher Policy: Reproduced under a Creative Commons License