Use of 13C315N1-serine or 13C315N1-methionine for studying methylation dynamics in cancer cell metabolism and epigenetics

Newman, A. C., Labuschagne, C. F., Vousden, K. H. and Maddocks, O. D.K. (2019) Use of 13C315N1-serine or 13C315N1-methionine for studying methylation dynamics in cancer cell metabolism and epigenetics. In: Haznadar, M. (ed.) Cancer Metabolism: Methods and Protocols. Series: Methods in molecular biology (1928). Humana Press ; Springer: New York, NY, pp. 55-67. ISBN 9781493990269 (doi:10.1007/978-1-4939-9027-6_4)

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Abstract

Tracing the fate of carbon-13 (C) labeled metabolites within cells by liquid chromatography mass spectrometry (LCMS) is a powerful analytical technique used for many years in the study of cell metabolism. Conventional experiments using LCMS and labeled nutrients tend to track the incorporation of C from exogenous nutrients (such as amino acids) into other, relatively proximal, cellular metabolites. Several labs have extended this technique to track transfer of C from the metabolite pool onto macromolecules, such as DNA, where methylation acts as an important functional modification. Here we describe a complete method that integrates previously established techniques to simultaneously track the use of C-serine or C-methionine into metabolite pools of the methionine cycle and into methylation of DNA and RNA. Given the ability to track methyl-transfer in a time-dependent way, this technique can provide temporal information about active methyl-transfer as well as quantification of total DNA/RNA methylation levels.

Item Type:Book Sections
Status:Published
Glasgow Author(s) Enlighten ID:Newman, Dr Alice and Vousden, Karen and Labuschagne, Dr Christiaan Fred and Maddocks, Dr Oliver
Authors: Newman, A. C., Labuschagne, C. F., Vousden, K. H., and Maddocks, O. D.K.
Subjects:Q Science > Q Science (General)
College/School:College of Medical Veterinary and Life Sciences > Institute of Cancer Sciences
Journal Name:Methods in molecular biology (Clifton, N.J.)
Publisher:Humana Press ; Springer
ISSN:1940-6029
ISBN:9781493990269
Published Online:07 February 2019

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