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Substrate specificity of the neutral sphingomyelinase from Trypanosoma brucei

Published online by Cambridge University Press:  05 November 2018

Emily A. Dickie ,
Simon A. Young  and
Terry K. Smith Open the ORCID record for Terry K. Smith [Opens in a new window]
Emily A. Dickie
Affiliation:
Biomedical Sciences Research Complex, Schools of Biology and Chemistry, University of St Andrews, Fife, KY16 9ST, UK
Simon A. Young
Affiliation:
Biomedical Sciences Research Complex, Schools of Biology and Chemistry, University of St Andrews, Fife, KY16 9ST, UK
Terry K. Smith*
Affiliation:
Biomedical Sciences Research Complex, Schools of Biology and Chemistry, University of St Andrews, Fife, KY16 9ST, UK
*
Author for correspondence: Terry K. Smith, E-mail: tks1@st-andrews.ac.uk
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Abstract

The kinetoplastid parasite Trypanosoma brucei causes African trypanosomiasis in both humans and animals. Infections place a significant health and economic burden on developing nations in sub-Saharan Africa, but few effective anti-parasitic treatments are currently available. Hence, there is an urgent need to identify new leads for drug development. The T. brucei neutral sphingomyelinase (TbnSMase) was previously established as essential to parasite survival, consequently being identified as a potential drug target. This enzyme may catalyse the single route to sphingolipid catabolism outside the T. brucei lysosome. To obtain new insight into parasite sphingolipid catabolism, the substrate specificity of TbnSMase was investigated using electrospray ionization tandem mass spectrometry (ESI-MS/MS). Recombinant TbnSMase was shown to degrade sphingomyelin, inositol-phosphoceramide and ethanolamine-phosphoceramide sphingolipid substrates, consistent with the sphingolipid complement of the parasites. TbnSMase also catabolized ceramide-1-phosphate, but was inactive towards sphingosine-1-phosphate. The broad-range specificity of this enzyme towards sphingolipid species is a unique feature of TbnSMase. Additionally, ESI-MS/MS analysis revealed previously uncharacterized activity towards lyso-phosphatidylcholine despite the enzyme's inability to degrade phosphatidylcholine. Collectively, these data underline the enzyme's importance in choline homoeostasis and the turnover of sphingolipids in T. brucei.

Keywords

Activity assaycholineenzymelipid catabolismlipid extractionmass spectrometrysphingolipid
Type
Research Article
Information
Parasitology , Volume 146 , Issue 5 , April 2019 , pp. 604 - 616
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Copyright
Copyright © Cambridge University Press 2018 

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Footnotes

*

Current address: Wellcome Trust Centre for Molecular Parasitology, Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, G12 8TA, UK.

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