title: Discovery of a chemical probe for the L3MBTL3 methyllysine reader domain.
abstract: We describe the discovery of UNC1215, a potent and selective chemical probe for the methyllysine (Kme) reading function of L3MBTL3, a member of the malignant brain tumor (MBT) family of chromatin-interacting transcriptional repressors. UNC1215 binds L3MBTL3 with a K(d) of 120 nM, competitively displacing mono- or dimethyllysine-containing peptides, and is greater than 50-fold more potent toward L3MBTL3 than other members of the MBT family while also demonstrating selectivity against more than 200 other reader domains examined. X-ray crystallography identified a unique 2:2 polyvalent mode of interaction between UNC1215 and L3MBTL3. In cells, UNC1215 is nontoxic and directly binds L3MBTL3 via the Kme-binding pocket of the MBT domains. UNC1215 increases the cellular mobility of GFP-L3MBTL3 fusion proteins, and point mutants that disrupt the Kme-binding function of GFP-L3MBTL3 phenocopy the effects of UNC1215 on localization. Finally, UNC1215 was used to reveal a new Kme-dependent interaction of L3MBTL3 with BCLAF1, a protein implicated in DNA damage repair and apoptosis.
authors.affiliation: Center for Integrative Chemical Biology and Drug Discovery, Division of Chemical Biology and Medicinal Chemistry, University of North Carolina Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
authors.forename: Lindsey I
authors.initials: LI
authors.lastname: James
authors.forename: Dalia
authors.initials: D
authors.lastname: Barsyte-Lovejoy
authors.forename: Nan
authors.initials: N
authors.lastname: Zhong
authors.forename: Liubov
authors.initials: L
authors.lastname: Krichevsky
authors.forename: Victoria K
authors.initials: VK
authors.lastname: Korboukh
authors.forename: J Martin
authors.initials: JM
authors.lastname: Herold
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: MacNevin
authors.forename: Jacqueline L
authors.initials: JL
authors.lastname: Norris
authors.forename: Cari A
authors.initials: CA
authors.lastname: Sagum
authors.forename: Wolfram
authors.initials: W
authors.lastname: Tempel
authors.forename: Edyta
authors.initials: E
authors.lastname: Marcon
authors.forename: Hongbo
authors.initials: H
authors.lastname: Guo
authors.forename: Cen
authors.initials: C
authors.lastname: Gao
authors.forename: Xi-Ping
authors.initials: XP
authors.lastname: Huang
authors.forename: Shili
authors.initials: S
authors.lastname: Duan
authors.forename: Andrew
authors.initials: A
authors.lastname: Emili
authors.forename: Jack F
authors.initials: JF
authors.lastname: Greenblatt
authors.forename: Dmitri B
authors.initials: DB
authors.lastname: Kireev
authors.forename: Jian
authors.initials: J
authors.lastname: Jin
authors.forename: William P
authors.initials: WP
authors.lastname: Janzen
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Brown
authors.forename: Mark T
authors.initials: MT
authors.lastname: Bedford
authors.forename: Cheryl H
authors.initials: CH
authors.lastname: Arrowsmith
authors.forename: Stephen V
authors.initials: SV
authors.lastname: Frye
mesh_terms: D001549:Benzamides / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D001667:Binding, Competitive / Q000187:drug effects; D018360:Crystallography, X-Ray; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D055808:Drug Discovery; D057809:HEK293 Cells; D006801:Humans; D008239:Lysine / Q000031:analogs & derivatives* / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D015335:Molecular Probes / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D015394:Molecular Structure; D010880:Piperidines / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D017434:Protein Structure, Tertiary; D012097:Repressor Proteins / Q000378:metabolism; D013329:Structure-Activity Relationship; D025521:Tumor Suppressor Proteins / Q000378:metabolism
chemical_list: C119386:BCLAF1 protein, human; D001549:Benzamides; D004268:DNA-Binding Proteins; C502190:L3MBTL3 protein, human; D015335:Molecular Probes; D010880:Piperidines; D012097:Repressor Proteins; D025521:Tumor Suppressor Proteins; C580998:UNC1215; C049907:2-methyllysine; D008239:Lysine