A General Molecular Affinity Strategy for Global Detection and Proteomic Analysis of Lysine Methylation

作者：Moore Kaitlyn E; Carlson Scott M; Camp Nathan D; Cheung Peggie; James Richard G; Chua Katrin F; Wolf Yadlin Alejandro; Gozani Or^*

来源：Molecular Cell, 2013, 50(3): 444-456.

DOI：10.1016/j.molcel.2013.03.005

Lysine methylation of histone proteins regulates chromatin dynamics and plays important roles in diverse physiological and pathological processes. However, beyond histone proteins, the proteome-wide extent of lysine methylation remains largely unknown. We have engineered the naturally occurring MBT domain repeats of L3MBTL1 to serve as a universal affinity reagent for detecting, enriching, and identifying proteins carrying a mono- or dimethylated lysine. The domain is broadly specific for methylated lysine ("pan-specific") and can be applied to any biological system. We have used our approach to demonstrate that SIRT1 is a substrate of the methyltransferase G9a both in vitro and in cells, to perform proteome-wide detection and enrichment of methylated proteins, and to identify candidate in-cell substrates of G9a and the related methyltransferase GLP. Together, our results demonstrate a powerful new approach for global and quantitative analysis of methylated lysine, and they represent the first systems biology understanding of lysine methylation.

• 出版日期2013-5-9