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Covalent inhibition of ERK docking interactions
Author(s) -
Kaoud Tamer Saad,
Johnson William H.,
Piserchio Andrea,
Ebelt Nancy D,
Warthaka Mangalika,
Cano Micael,
Sammons Rachel,
Wang Qiantao,
Ren Pengyu,
Ghose Ranajeet,
Dalby Kevin N.
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.856.11
Subject(s) - mapk/erk pathway , phosphorylation , kinase , docking (animal) , chemistry , hek 293 cells , microbiology and biotechnology , protein kinase a , biochemistry , mutant , biology , receptor , medicine , nursing , gene
The extracellular‐regulated kinase (ERK) signaling pathway has been implicated in the regulation of several cellular functions, including cell differentiation, proliferation, and inflammatory responses. Upon stimulation, ERKs have been reported to phosphorylate a large number of substrates localized either in the cytoplasm or the nucleus. Taking advantage of our understanding of ERK docking interactions, we tried to design/discover substrate‐selective inhibitors that target the protein‐binding site of ERK. Here we report the discovery of the first irreversible ERK inhibitor that targets its protein‐docking site. Protein NMR, mass spectrometry, mutagenesis and molecular docking studies showed that the compound forms a covalent bond with a conserved cysteine residue C‐159. Extensive biochemical studies have been employed to estimate its kinetic parameters and its kinase‐selectivity profile. Finally, the new ERK inhibitor was shown to inhibit both ERK phosphorylation and its ability to phosphorylate downstream substrates (e.g. p90RSK and Elk‐1) in HEK293 cells. The inhibitor showed dose‐dependent inhibitory effects on HEK293 and melanoma A375 cell proliferation and viability. Targeting ERK in cells was confirmed using a chemical‐genetic approach where the C159A mutant was used to rescue the effects of this compound on ERK signaling. The discovered compound represents a new class of ERK inhibitor that uniqely labels its protein binding site. Further development may result in a pharmacologically useful inhibitor. Support or Funding Information This research has been supported by the following grants from the National Institutes of Health: GM059802 (to K.N.D.),GM084278 (to R.G.), and G12 MD007603 (towards partial support of the NMR facilities at The City College of New York). R.G. and K.D. are members of the New York Structural Biology Center, NYSTAR facility. T.S.K. acknowledges Cancer Prevention and Research Institute of Texas Postdoctoral Training Award (RP140108).