title: Preparation, Biochemical Analysis, and Structure Determination of SET Domain Histone Methyltransferases.
abstract: In eukaryotes, several lysine residues on histone proteins are methylated. This posttranslational modification is linked to a myriad of nuclear-based transactions such as epigenetic inheritance of heterochromatin, regulation of gene expression, DNA damage repair, and DNA replication. The majority of the enzymes responsible for writing these marks onto chromatin belong to the SET domain family of histone lysine methyltransferases. Although they often share important structural features, including a conserved catalytic domain, SET domain enzymes use different mechanisms to achieve substrate recognition, mono-, di-, or trimethylate lysine residues and some require other proteins to achieve maximal methyltransferase activity. In this chapter, we summarize our efforts to purify, crystallize, and enzymatically characterize SET domain enzymes with a specific focus on the histone H3K27 monomethyltransferase ATXR5.
authors.affiliation: Ottawa Institute of Systems Biology, University of Ottawa, Ottawa, ON, Canada.
authors.forename: E
authors.initials: E
authors.lastname: Bergamin
authors.affiliation: Ottawa Institute of Systems Biology, University of Ottawa, Ottawa, ON, Canada. Electronic address: jean-francois.couture@uottawa.ca.
authors.forename: J F
authors.initials: JF
authors.lastname: Couture
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D018360:Crystallography, X-Ray / Q000379:methods; D057075:Enzyme Assays / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D011495:Histone-Lysine N-Methyltransferase / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D008958:Models, Molecular; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D000072417:Protein Domains; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012278:Ricinus / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D015854:Up-Regulation
chemical_list: D010940:Plant Proteins; D011994:Recombinant Proteins; D011495:Histone-Lysine N-Methyltransferase
keywords: ATXR5; Histone H3K27 monomethylation; Histone lysine methylation; Methyltransferase assays; SET domain crystallization; SET domain enzyme; SET domain overexpression; SET domain purification; Structure-based drug development