An affinity-based probe for methyltransferase enzymes based on sinefungin

作者：Lafreniere Matthew A; Desrochers Genevieve F; Mekbib Kedous; Pezacki John Paul^*

来源：Canadian Journal of Chemistry, 2017, 95(10): 1059-1063.

DOI：10.1139/cjc-2017-0168

• affinity-based protein profiling
• sinefungin
• enzyme activity
• methyltransferases
• epigenetics

Epigenetics control numerous cellular processes such as gene transcription, signal transduction, and protein stabilization. An understanding of epigenetic mechanisms can lead to the development of therapeutic agents for various diseases. Herein, we report the design and synthesis of a sinefungin affinity-probe (BpyneSF) that targets methyltranferase enzymes and proteins involved in recognition of methylation. This probe contains a bioorthogonal alkyne residue for conjugation using the copper-catalyzed azide-alkyne cycloaddition and a photoactivatable crosslinker group for covalent attachment of the probe to its proteomic targets. We investigate the efficiency and selectivity of the probe to inhibit and label methyltransferase enzymes, and we demonstrate, through in-gel fluorescence, on-bead digestion, and tandem mass spectrometry, that BpyneSF can label methyltransferase SETD2 and reader proteins in vitro. These results establish the utility of BpyneSF as a tool for affinity-based protein profiling in complex biological environments.

• 出版日期2017-10