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)

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: Labuschagne, Dr Christiaan Fred and Maddocks, Professor Oliver and Newman, Dr Alice and Vousden, Karen
• 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 > School 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