Approaches and Guidelines for the Identification of Novel Substrates of Protein Lysine Methyltransferases. Issue 9 (22nd September 2016)

• Record Type: Journal Article
• Authors: Kudithipudi, Srikanth; Jeltsch, Albert
• Is Part Of: Cell chemical biology. Volume 23:Issue 9(2016)
• Page Start: 1049
• Page End: 1055
• Publication Date: 2016-09-22
• Subjects: lysine methylation -- protein lysine methyltransferase -- post-translational modification -- enzyme specificity -- proteome; Biochemistry -- Periodicals; 572.05
• DOI: 10.1016/j.chembiol.2016.07.013
• Languages: English
• ISSNs: 2451-9456

Abstract

Abstract : Protein lysine methylation is emerging as a general post-translational modification (PTM) with essential functions regulating protein stability, activity, and protein-protein interactions. One of the outstanding challenges in this field is linking protein lysine methyltransferases (PKMTs) with specific substrates and lysine methylation events in a systematic manner. Inability to validate reported PKMT substrates delayed progress in the field and cast unnecessary doubt about protein lysine methylation as a truly general PTM. Here, we aim to provide a concise guide to help avoid some of the most common pitfalls in studies searching for new PKMT substrates and propose a set of seven basic biochemical rules: (1) include positive controls; (2) use target lysine mutations of substrate proteins as negative controls; (3) use inactive enzyme variants as negative controls; (4) report quantitative methylation data; (5) consider PKMT specificity; (6) validate methyl lysine antibodies; and (7) connect cellular and in vitro results. We explain the logic behind them and discuss how they should be implemented in the experimental work. Abstract : Protein lysine methylation is a post-translational modification mediated by protein lysine methyltransferases (PKMTs). Although the biological significance of lysine methylation is well established, validating PKMT substrates has proven to be a challenge. Kudithipudi and Jeltsch build on their experience to propose a set of biochemical guidelines and best practices to help address this bottleneck and to better understand the biology of protein lysine methylation.