Determining the Mode of Action of Antimalarial Drugs Using Time-Resolved LC-MS-Based Metabolite Profiling View Full Text


Ontology type: schema:Chapter     


Chapter Info

DATE

2018-11-13

AUTHORS

Simon A. Cobbold , Malcolm J. McConville

ABSTRACT

Methods for assessing the mode of action of new antimalarial compounds identified in high throughput phenotypic screens are needed to triage and facilitate lead compound development and to anticipate potential resistance mechanisms that might emerge. Here we describe a mass spectrometry-based approach for detecting metabolic changes in asexual erythrocytic stages of Plasmodium falciparum induced by antimalarial compounds. Time-resolved or concentration-resolved measurements are used to discriminate between putative targets of the compound and nonspecific and/or downstream secondary metabolic effects. These protocols can also be coupled with 13C-stable-isotope tracing experiments under nonequilibrative (or nonstationary) conditions to measure metabolic dynamics following drug exposure. Time-resolved 13C-labeling studies greatly increase confidence in target assignment and provide a more comprehensive understanding of the metabolic perturbations induced by small molecule inhibitors. The protocol provides details on the experimental design, Plasmodium falciparum culture, sample preparation, analytical approaches, and data analysis used in either targeted (pathway focused) or untargeted (all detected metabolites) analysis of drug-induced metabolic perturbations. More... »

PAGES

225-239

Book

TITLE

Microbial Metabolomics

ISBN

978-1-4939-8756-6
978-1-4939-8757-3

Author Affiliations

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/978-1-4939-8757-3_12

DOI

http://dx.doi.org/10.1007/978-1-4939-8757-3_12

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1109831530

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/30421232


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