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Updated: Sep 12, 2025

Efficient Purification and LC-MS/MS-based Assay Development for Ten-Eleven Translocation-2 5-Methylcytosine Dioxygenase
Published on: October 15, 2018
Protocol for elucidating metabolite binding and regulation of TET2 dioxygenase
Shuyuan Zhang1, Zhou-Li Cheng2, Dan Ye1
1Huashan Hospital, Fudan University, and International Co-laboratory of Medical Epigenetics and Metabolism of Ministry of Science and Technology, and Key Laboratory of Metabolism and Molecular Medicine of Ministry of Education, and Molecular and Cell Biology Lab, Institutes of Biomedical Sciences, Shanghai Medical College of Fudan University, Shanghai 200032, China.
Abstract:
Epigenetic enzyme activity is coupled to cellular metabolism through their reliance on metabolic cofactors and substrates. Here, we describe steps for combining biochemical assays and saturation transfer difference (STD) NMR spectroscopy to experimentally validate metabolite binding and assess the effect on TET2 activity. This protocol enables the identification of both TET2 activators and inhibitors, providing a framework for studying the interplay between metabolism and epigenetic regulation. For complete details on the use and execution of this protocol, please refer to Cheng et al.1.
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