title: A general molecular affinity strategy for global detection and proteomic analysis of lysine methylation.
abstract: 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.
authors.affiliation: Department of Biology, Stanford University, Stanford, CA 94305, USA.
authors.forename: Kaitlyn E
authors.initials: KE
authors.lastname: Moore
authors.forename: Scott M
authors.initials: SM
authors.lastname: Carlson
authors.forename: Nathan D
authors.initials: ND
authors.lastname: Camp
authors.forename: Peggie
authors.initials: P
authors.lastname: Cheung
authors.forename: Richard G
authors.initials: RG
authors.lastname: James
authors.forename: Katrin F
authors.initials: KF
authors.lastname: Chua
authors.forename: Alejandro
authors.initials: A
authors.lastname: Wolf-Yadlin
authors.forename: Or
authors.initials: O
authors.lastname: Gozani
mesh_terms: D000818:Animals; D002460:Cell Line; D057809:HEK293 Cells; D006801:Humans; D007313:Insecta; D008239:Lysine / Q000235:genetics* / Q000378:metabolism*; D008745:Methylation; D017434:Protein Structure, Tertiary; D020543:Proteome / Q000235:genetics* / Q000378:metabolism*; D040901:Proteomics / Q000379:methods; D061987:Sf9 Cells; D056564:Sirtuin 1 / Q000235:genetics / Q000378:metabolism
chemical_list: D020543:Proteome; C447939:SIRT1 protein, human; D056564:Sirtuin 1; D008239:Lysine