title: Identification of Inhibitors of the Association of ZAP-70 with the T Cell Receptor by High-Throughput Screen.
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.
authors.affiliation: 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.|Plexxikon Inc., Berkeley, CA, USA.
authors.forename: Patrick R
authors.initials: PR
authors.lastname: Visperas
authors.affiliation: 3 Department of Pharmaceutical Chemistry, Small Molecule Discovery Center, University of California, San Francisco, CA, USA.
authors.forename: Christopher G
authors.initials: CG
authors.lastname: Wilson
authors.affiliation: 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.|Omniox Inc., San Carlos, CA, USA.
authors.forename: Jonathan A
authors.initials: JA
authors.lastname: Winger
authors.affiliation: 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.|Janssen Pharmaceuticals Inc., Titusville, NJ, USA.
authors.forename: Qingrong
authors.initials: Q
authors.lastname: Yan
authors.affiliation: 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.
authors.forename: Kevin
authors.initials: K
authors.lastname: Lin
authors.affiliation: 3 Department of Pharmaceutical Chemistry, Small Molecule Discovery Center, University of California, San Francisco, CA, USA.
authors.forename: Michelle R
authors.initials: MR
authors.lastname: Arkin
authors.affiliation: 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.
authors.forename: Arthur
authors.initials: A
authors.lastname: Weiss
authors.affiliation: 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 Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.
authors.forename: John
authors.initials: J
authors.lastname: Kuriyan
mesh_terms: D002474:Cell-Free System / Q000737:chemistry; D005454:Fluorescence Polarization / Q000379:methods; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods; D015870:Gene Expression; D057166:High-Throughput Screening Assays; D006801:Humans; D007155:Immunologic Factors / Q000737:chemistry / Q000494:pharmacology*; D010455:Peptides / Q000037:antagonists & inhibitors / Q000235:genetics / Q000276:immunology; D010766:Phosphorylation / Q000187:drug effects; D011485:Protein Binding / Q000187:drug effects; D047428:Protein Kinase Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D011948:Receptors, Antigen, T-Cell / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000276:immunology; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D015398:Signal Transduction; D054852:Small Molecule Libraries / Q000737:chemistry / Q000494:pharmacology*; D051746:ZAP-70 Protein-Tyrosine Kinase / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000276:immunology
chemical_list: D007155:Immunologic Factors; D010455:Peptides; D047428:Protein Kinase Inhibitors; D011948:Receptors, Antigen, T-Cell; D011994:Recombinant Proteins; D054852:Small Molecule Libraries; D051746:ZAP-70 Protein-Tyrosine Kinase; C497261:ZAP70 protein, human
keywords: T cell; ZAP-70; covalent inhibitor; fluorescence polarization (FP); kinase; time-resolved fluorescence resonance energy transfer (TR-FRET)