Use of 13C315N1-serine or 13C515N1-methionine for studying methylation dynamics in cancer cell metabolism and epigenetics

Journal Article: Methods in Molecular BiologyYear Published: (2019) Volume Number: 1928, Article Number: 55-67

Authors

Newman,Alice C; Labuschagne,Christiaan F; Vousden,Karen H; Maddocks,Oliver DK

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.