Supplementary Materials

Supplementary Materials for:

The catalytic activity of the kinase ZAP-70 mediates basal signaling and negative feedback of the T cell receptor pathway

Hanna Sjölin-Goodfellow, Maria P. Frushicheva, Qinqin Ji, Debra A Cheng, Theresa A. Kadlecek, Aaron J. Cantor, John Kuriyan, Arup K. Chakraborty,* Arthur R. Salomon,* Arthur Weiss*

*Corresponding author. E-mail: as{at}brown.edu (A.R.S.); arupc{at}mit.edu (A.K.C.); aweiss{at}medicine.ucsf.edu (A.W.)

This PDF file includes:

  • Experimental procedures.
  • Computational modeling.
  • Fig. S1. Reproducibility of proteomic quantitative data in the absence of TCR stimulation.
  • Fig. S2. Reproducibility of proteomic quantitative data for all TCR stimulation time points.
  • Fig. S3. Validation of inhibitor specificity.
  • Fig. S4. Binding of the SH2 domain of Lck to monophosphorylated TCR ζ-chain ITAM peptides.
  • Fig. S5. The number of rebinding events between enzymes and substrates upon variation of different kinetic parameters.
  • Fig. S6. Description of the ZAP-70 allosteric model and the results of calculations with this model.
  • Fig. S7. Sensitivity analysis of the kinetic parameters used in calculations for the “ITAM and Lck” model.
  • Fig. S8. Sensitivity analysis of the kinetic parameters used in calculations for the “ITAMs” model.
  • Fig. S9. Sensitivity analysis of the kinetic parameters used in calculations for the ZAP-70 allosteric model (part I).
  • Fig. S10. Sensitivity analysis of the kinetic parameters used in calculations for the ZAP-70 allosteric model (part II).
  • Legends for tables S1 and S2
  • Table S3. Fold change between HXJ-42–treated and DMSO-treated ZAP-70AS cells and Q values for peptides listed in Table 1.
  • Table S4. Binding parameters for the Lck SH2 domain and monophosphorylated ζ ITAM peptides determined by isothermal titration calorimetry at 25°C.
  • Table S5. Concentrations of species used in calculations for all models (volume = 1 μm3).
  • Table S6. Reactions and kinetic parameters used in calculations for the ZAP-70–mediated negative feedback model and models used to reproduce an asymmetry in ITAM phosphorylation.
  • Table S7. Reactions and kinetic parameters used in calculations for the ZAP-70 allosteric function model.
  • Table S8. Sensitivity analysis of the concentrations of signaling molecules used in calculations for the “ITAM and Lck” and “ITAMs” models.
  • Table S9. Sensitivity analysis of the kinetic parameters used in calculations for the “ITAM and Lck” model.
  • Table S10. Sensitivity analysis of the kinetic parameters used in calculations for the “ITAMs” model.
  • Table S11. Sensitivity analysis of the concentrations of signaling molecules used in calculations for the ZAP-70 allosteric model.
  • Table S12. Sensitivity analysis of the kinetic parameters used in calculations for ZAP-70 allosteric model.
  • References (7994)

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Other Supplementary Material for this manuscript includes the following:

  • Table S1 (Microsoft Excel format). Complete list of sequence and phosphorylation site assignments of all identified phosphopeptides with corresponding SIC peak areas and statistics, protein association numbers, gene ontology, and KEGG functional annotation.
  • Table S2 (Microsoft Excel format). Complete list of phosphopeptides detected from every replicate and time point of TCR stimulation.

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Citation: H. Sjölin-Goodfellow, M. P. Frushicheva, Q. Ji, D. A. Cheng, T. A. Kadlecek, A. J. Cantor, J. Kuriyan, A. K. Chakraborty, A. R. Salomon, A. Weiss, The catalytic activity of the kinase ZAP-70 mediates basal signaling and negative feedback of the T cell receptor pathway. Sci. Signal. 8, ra49 (2015).

© 2015 American Association for the Advancement of Science