title: A rationally-designed chimeric KDM1A/KDM1B histone demethylase tower domain deletion mutant retaining enzymatic activity.
abstract: A target with therapeutic potential, lysine-specific demethylase 1A (KDM1A) is a regulator of gene expression whose tower domain is a protein-protein interaction motif. This domain facilitates the interaction of KDM1A with coregulators and multiprotein complexes that direct its activity to nucleosomes. We describe the design and characterization of a chimeric 'towerless' KDM1A, termed nΔ150 KDM1AΔTower KDM1B chimera (chKDM1AΔTower), which incorporates a region from the paralog lysine-specific demethylase 1B (KDM1B). This chimera copurifies with FAD and displays demethylase activity, but fails to bind the partner protein corepressor of the RE1-silencing transcription factor (CoREST). We conclude that KDM1A catalysis can be decoupled from tower-dependent interactions, lending chKDM1AΔTower useful for dissecting molecular contributions to KDM1A function.
authors.affiliation: Department of Chemistry, Duke University, Durham, NC, USA.
authors.forename: Jonathan M
authors.initials: JM
authors.lastname: Burg
authors.affiliation: Trinity College of Arts & Sciences, Duke University, Durham, NC, USA.
authors.forename: Alan T
authors.initials: AT
authors.lastname: Makhoul
authors.affiliation: Duke University Human Vaccine Institute Macromolecular Crystallography Center, Duke University School of Medicine, Durham, NC, USA.
authors.forename: Charles W
authors.initials: CW
authors.lastname: Pemble
authors.affiliation: Department of Chemistry, Duke University, Durham, NC, USA.
authors.forename: Jennifer E
authors.initials: JE
authors.lastname: Link
authors.affiliation: Trinity College of Arts & Sciences, Duke University, Durham, NC, USA.
authors.forename: Frederick J
authors.initials: FJ
authors.lastname: Heller
authors.affiliation: Department of Chemistry, Duke University, Durham, NC, USA. Electronic address: dewey.mccafferty@duke.edu.
authors.forename: Dewey G
authors.initials: DG
authors.lastname: McCafferty
mesh_terms: D000595:Amino Acid Sequence; D056466:Histone Demethylases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D015202:Protein Engineering; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D017384:Sequence Deletion
chemical_list: D011993:Recombinant Fusion Proteins; D056466:Histone Demethylases; C495195:KDM1A protein, human
keywords: Chimera; CoREST; Deletion mutant; Enzyme engineering; KDM1A/LSD1; Tower domain