A Chemical Probe Strategy for Interrogating Inhibitor Selectivity Across the MEK Kinase Family

Kristine K. Deibler, Rama K. Mishra, Matthew R. Clutter, Aleksandar Antanasijevic, Raymond Bergan, Michael Caffrey, Karl A. Scheidt

Research output: Research - peer-reviewArticle

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Abstract

MEK4 is an upstream kinase in MAPK signaling pathways where it phosphorylates p38 MAPK and JNK in response to mitogenic and cellular stress queues. MEK4 is overexpressed and induces metastasis in advanced prostate cancer lesions. However, the value of MEK4 as an oncology target has not been pharmacologically validated because selective chemical probes targeting MEK4 have not been developed. Despite a high level of sequence homology in the ATP-binding site, most reported MEK inhibitors are selective for MEK1/2 and display reduced potency toward other MEKs. Here, we present the first functional and binding selectivity-profiling platform of the MEK family. We applied the platform to profile a set of known kinase inhibitors and used the results to develop an in silico approach for small molecule docking against MEK proteins. The docking studies identified molecular features of the ligands and corresponding amino acids in MEK proteins responsible for high affinity binding versus those driving selectivity. WaterLOGSY and saturation transfer difference (STD) NMR spectroscopy techniques were utilized to understand the binding modes of active compounds. Further minor synthetic manipulations provide a proof of concept by showing how information gained through this platform can be utilized to perturb selectivity across the MEK family. This inhibitor-based approach pinpoints key features governing MEK family selectivity and clarifies empirical selectivity profiles for a set of kinase inhibitors. Going forward, the platform provides a rationale for facilitating the development of MEK-selective inhibitors, particularly MEK4 selective inhibitors, and repurposing of kinase inhibitors for probing the structural selectivity of isoforms.

LanguageEnglish (US)
Pages1245-1256
Number of pages12
JournalACS Chemical Biology
Volume12
Issue number5
DOIs
StatePublished - May 19 2017

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MAP Kinase Kinase Kinases
Mitogen-Activated Protein Kinase Kinases
Phosphotransferases
Proteins
p38 Mitogen-Activated Protein Kinases
Sequence Homology
Computer Simulation
Prostatic Neoplasms
Protein Isoforms
Magnetic Resonance Spectroscopy
Adenosine Triphosphate
Binding Sites
Neoplasm Metastasis
Ligands
Amino Acids
mitogen-activated protein kinase kinase kinase 12
Oncology
Nuclear magnetic resonance spectroscopy
Display devices
Molecules

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine

Cite this

Deibler, K. K., Mishra, R. K., Clutter, M. R., Antanasijevic, A., Bergan, R., Caffrey, M., & Scheidt, K. A. (2017). A Chemical Probe Strategy for Interrogating Inhibitor Selectivity Across the MEK Kinase Family. ACS Chemical Biology, 12(5), 1245-1256. DOI: 10.1021/acschembio.6b01060

A Chemical Probe Strategy for Interrogating Inhibitor Selectivity Across the MEK Kinase Family. / Deibler, Kristine K.; Mishra, Rama K.; Clutter, Matthew R.; Antanasijevic, Aleksandar; Bergan, Raymond; Caffrey, Michael; Scheidt, Karl A.

In: ACS Chemical Biology, Vol. 12, No. 5, 19.05.2017, p. 1245-1256.

Research output: Research - peer-reviewArticle

Deibler, KK, Mishra, RK, Clutter, MR, Antanasijevic, A, Bergan, R, Caffrey, M & Scheidt, KA 2017, 'A Chemical Probe Strategy for Interrogating Inhibitor Selectivity Across the MEK Kinase Family' ACS Chemical Biology, vol 12, no. 5, pp. 1245-1256. DOI: 10.1021/acschembio.6b01060
Deibler KK, Mishra RK, Clutter MR, Antanasijevic A, Bergan R, Caffrey M et al. A Chemical Probe Strategy for Interrogating Inhibitor Selectivity Across the MEK Kinase Family. ACS Chemical Biology. 2017 May 19;12(5):1245-1256. Available from, DOI: 10.1021/acschembio.6b01060
Deibler, Kristine K. ; Mishra, Rama K. ; Clutter, Matthew R. ; Antanasijevic, Aleksandar ; Bergan, Raymond ; Caffrey, Michael ; Scheidt, Karl A./ A Chemical Probe Strategy for Interrogating Inhibitor Selectivity Across the MEK Kinase Family. In: ACS Chemical Biology. 2017 ; Vol. 12, No. 5. pp. 1245-1256
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