Identification of Inhibitors of the Association of ZAP-70 with the T Cell Receptor by High-Throughput Screen

  • SLAS Discov. 2017 Mar;22(3):324-331. doi: 10.1177/1087057116681407.
Patrick R Visperas  1  2 ,  Christopher G Wilson  3 ,  Jonathan A Winger  1  4 ,  Qingrong Yan  1  5 ,  Kevin Lin  1 ,  Michelle R Arkin  3 ,  Arthur Weiss  6 ,  John Kuriyan  1  7
Affiliations
  • 1. 1 Department of Molecular and Cell Biology and Department of Chemistry, California Institute of Quantitative Biosciences and Howard Hughes Medical Institute, University of California, Berkeley, CA, USA.
  • 2. Plexxikon Inc., Berkeley, CA, USA.
  • 3. 3 Department of Pharmaceutical Chemistry, Small Molecule Discovery Center, University of California, San Francisco, CA, USA.
  • 4. Omniox Inc., San Carlos, CA, USA.
  • 5. Janssen Pharmaceuticals Inc., Titusville, NJ, USA.
  • 6. 4 Department of Medicine, Rosalind Russell and Ephrain P. Engleman Rheumatology Research Center for Arthritis and Howard Hughes Medical Institute, University of California, San Francisco, CA, USA.
  • 7. 2 Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.
PMID: 27932698 DOI: 10.1177/1087057116681407
Abstract

ZAP-70 is a critical molecule in the transduction of T cell antigen receptor signaling and the activation of T cells. Upon activation of the T cell antigen receptor, ZAP-70 is recruited to the intracellular ζ-chains of the T cell receptor, where ZAP-70 is activated and colocalized with its substrates. Inhibitors of ZAP-70 could potentially function as treatments for autoimmune diseases or organ transplantation. In this work, we present the design, optimization, and implementation of a screen for inhibitors that would disrupt the interaction between ZAP-70 and the T cell antigen receptor. The screen is based on a fluorescence polarization assay for peptide binding to ZAP-70.

Keywords
T cell; ZAP-70; covalent inhibitor; fluorescence polarization (FP); kinase; time-resolved fluorescence resonance energy transfer (TR-FRET).
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