Title: Protein Methylation Characterization Using NMR Without Isotopic Labeling
Abstract: Protein methylation is crucial in epigenetics, and targeting the involved methyltransferases shows great potential for therapeutic intervention with several inhibitors in clinical trials for oncology indications. Therefore, characterization of protein methylation is essential for understanding the methyltransferase function and discovering chemical inhibitors and antagonists. While NMR has been used to measure methylation rates, isotopic labeling of protein or methyl donors can be costly and cannot characterize demethylation of proteins extracted from natural sources. Our method employs a four-quantum filter 1H-13C experiment that selectively detects methyl groups, providing a simple way to characterize methylation and demethylation features of methyltransferases and demethylases, respectively, without requiring isotopic labeling. In our experiments, we successfully observed the methylation of H3 under lysates from various cells and tissues from mice with cancerous growth, showing that H3 is mono- and dimethylated in all the tested lysates, but at varying rates and levels. This method could aid in discovering chemical inhibitors and antagonists against methyltransferases related to diseases by characterizing the methylation features of suspected tumors or areas of concern.
Publication Year: 2023
Publication Date: 2023-01-01
Language: en
Type: preprint
Indexed In: ['crossref']
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