2. Academic Validation
  3. Discovery of a chemical probe for the L3MBTL3 methyllysine reader domain

Discovery of a chemical probe for the L3MBTL3 methyllysine reader domain

  • Nat Chem Biol. 2013 Mar;9(3):184-91. doi: 10.1038/nchembio.1157.
Lindsey I James 1,  Dalia Barsyte-Lovejoy,  Nan Zhong,  Liubov Krichevsky,  Victoria K Korboukh,  J Martin Herold,  Christopher J MacNevin,  Jacqueline L Norris,  Cari A Sagum,  Wolfram Tempel,  Edyta Marcon,  Hongbo Guo,  Cen Gao,  Xi-Ping Huang,  Shili Duan,  Andrew Emili,  Jack F Greenblatt,  Dmitri B Kireev,  Jian Jin,  William P Janzen,  Peter J Brown,  Mark T Bedford,  Cheryl H Arrowsmith,  Stephen V Frye
Affiliations

Affiliation

  • 1 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.
PMID: 23292653 DOI: 10.1038/nchembio.1157
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.

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