Analysis of cell metabolism using LC-MS and isotope tracers

Mackay, G. M., Zheng, L., Van Den Broek, N. J.F. and Gottlieb, E. (2015) Analysis of cell metabolism using LC-MS and isotope tracers. Methods in Enzymology, 561, pp. 171-196. (doi: 10.1016/bs.mie.2015.05.016) (PMID:26358905)

Full text not currently available from Enlighten.

Abstract

Here we discuss our methods to analyze small polar compounds involved in central carbon metabolism using LC-MS. Methods described include sample extraction procedures for cells and medium, as well as for plasma/serum, urine, CSF, and tissue samples. Different extraction solvents are assessed. Our methods for using 13C stable isotope tracers to examine the kinetics and distributions of mass isotopologues of many metabolites are discussed. Quantification methods are described for 13C stable isotope tracer experiments as well as for unlabeled experiments. These methods were applied in a fumarate hydratase deficient cell model to show how isotope tracing can demonstrate shifts in metabolic pathways and, together with metabolite exchange rates, can be used to gain insights into changes in cell metabolism.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Van Den Broek, Mr Niels and Zheng, Mr Liang and Gottlieb, Professor Eyal and Mackay, Dr Gillian
Authors: Mackay, G. M., Zheng, L., Van Den Broek, N. J.F., and Gottlieb, E.
College/School:College of Medical Veterinary and Life Sciences > School of Cancer Sciences
Journal Name:Methods in Enzymology
Publisher:Elsevier
ISSN:0076-6879
ISSN (Online):1557-7988
Published Online:17 August 2015

University Staff: Request a correction | Enlighten Editors: Update this record