Evaluation of Jumonji C lysine demethylase substrate preference to guide identification of in vitro substrates

Matthew Hoekstra; Anand Chopra; William G. Willmore; Kyle K. Biggar

STAR Protocols (Jun 2022)

Evaluation of Jumonji C lysine demethylase substrate preference to guide identification of in vitro substrates

Matthew Hoekstra,
Anand Chopra,
William G. Willmore,
Kyle K. Biggar

Affiliations

Matthew Hoekstra: Institute of Biochemistry and Department of Biology, Carleton University, 1125 Colonel By Drive, Ottawa, ON K1S 5B6, Canada
Anand Chopra: Institute of Biochemistry and Department of Biology, Carleton University, 1125 Colonel By Drive, Ottawa, ON K1S 5B6, Canada
William G. Willmore: Institute of Biochemistry and Department of Biology, Carleton University, 1125 Colonel By Drive, Ottawa, ON K1S 5B6, Canada
Kyle K. Biggar: Institute of Biochemistry and Department of Biology, Carleton University, 1125 Colonel By Drive, Ottawa, ON K1S 5B6, Canada; Corresponding author

Journal volume & issue: Vol. 3, no. 2
p. 101271

Abstract

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Summary: Within the realm of lysine methylation, the discovery of lysine methyltransferase (KMTs) substrates has been burgeoning because of established systematic substrate screening protocols. Here, we describe a protocol enabling the systematic identification of JmjC KDM substrate preference and in vitro substrates. Systematically designed peptide libraries containing methylated lysine residues are used to characterize enzyme-substrate preference and identify new candidate substrates in vitro.For complete details on the use and execution of this protocol, please refer to Hoekstra and Biggar (2021).

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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