title: An Integrative Proteomic Approach Identifies Novel Cellular SMYD2 Substrates.
abstract: Protein methylation is a post-translational modification with important roles in transcriptional regulation and other biological processes, but the enzyme-substrate relationship between the 68 known human protein methyltransferases and the thousands of reported methylation sites is poorly understood. Here, we propose a bioinformatic approach that integrates structural, biochemical, cellular, and proteomic data to identify novel cellular substrates of the lysine methyltransferase SMYD2. Of the 14 novel putative SMYD2 substrates identified by our approach, six were confirmed in cells by immunoprecipitation: MAPT, CCAR2, EEF2, NCOA3, STUB1, and UTP14A. Treatment with the selective SMYD2 inhibitor BAY-598 abrogated the methylation signal, indicating that methylation of these novel substrates was dependent on the catalytic activity of the enzyme. We believe that our integrative approach can be applied to other protein lysine methyltransferases, and help understand how lysine methylation participates in wider signaling processes.
authors.affiliation: Structural Genomics Consortium, University of Toronto , 101 College Street, MaRS Centre, South Tower, Toronto, Ontario M5G 1L7, Canada.
authors.forename: Hazem
authors.initials: H
authors.lastname: Ahmed
authors.affiliation: Princess Margaret Cancer Centre and Department of Medical Biophysics, University of Toronto , Toronto, Ontario M5S 1A8, Canada.
authors.forename: Shili
authors.initials: S
authors.lastname: Duan
authors.affiliation: Structural Genomics Consortium, University of Toronto , 101 College Street, MaRS Centre, South Tower, Toronto, Ontario M5G 1L7, Canada.|Princess Margaret Cancer Centre and Department of Medical Biophysics, University of Toronto , Toronto, Ontario M5S 1A8, Canada.
authors.forename: Cheryl H
authors.initials: CH
authors.lastname: Arrowsmith
authors.affiliation: Structural Genomics Consortium, University of Toronto , 101 College Street, MaRS Centre, South Tower, Toronto, Ontario M5G 1L7, Canada.
authors.forename: Dalia
authors.initials: D
authors.lastname: Barsyte-Lovejoy
authors.affiliation: Structural Genomics Consortium, University of Toronto , 101 College Street, MaRS Centre, South Tower, Toronto, Ontario M5G 1L7, Canada.|Department of Pharmacology and Toxicology, University of Toronto ; 1 King's College Circle, Toronto, Ontario M5S 1A8, Canada.
authors.forename: Matthieu
authors.initials: M
authors.lastname: Schapira
mesh_terms: D002460:Cell Line; D019295:Computational Biology; D011495:Histone-Lysine N-Methyltransferase / Q000378:metabolism*; D006801:Humans; D047468:Immunoprecipitation; D008745:Methylation; D011499:Protein Processing, Post-Translational; D040901:Proteomics / Q000379:methods*; D013379:Substrate Specificity
chemical_list: D011495:Histone-Lysine N-Methyltransferase; C512569:SMYD2 protein, human
keywords: SMYD2; data integration; lysine methylation; non-histone substrates; pan-methyl lysine antibody