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

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...

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Bibliographic Details
Published in:Molecular cell 2013-05, Vol.50 (3), p.444-456
Main Authors: Moore, Kaitlyn E., Carlson, Scott M., Camp, Nathan D., Cheung, Peggie, James, Richard G., Chua, Katrin F., Wolf-Yadlin, Alejandro, Gozani, Or
Format: Article
Language:English
Subjects:
Animals
Cell Line
chromatin
HEK293 Cells
histones
Humans
Insecta
lysine
Lysine - genetics
Lysine - metabolism
Methylation
methyltransferases
Protein Structure, Tertiary
Proteome - genetics
Proteome - metabolism
proteomics
Proteomics - methods
quantitative analysis
Sf9 Cells
Sirtuin 1 - genetics
Sirtuin 1 - metabolism
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Summary: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. ► 3xMBT domain is a “pan-specific” tool for detecting mono- and dimethylated lysine ► 3xMBT allows for rapid analysis of methylated proteins in vitro and in vivo ► Proteomic analysis shows that lysine methylation occurs widely across the proteome ► A cell-based proteomic assay allows global analysis of dynamic lysine methylation
ISSN:1097-2765
1097-4164
DOI:10.1016/j.molcel.2013.03.005