Supplementary Materials

Supplementary Materials for:

Instructive roles for cytokine-receptor binding parameters in determining signaling and functional potency

Ignacio Moraga, David Richter, Stephan Wilmes, Hauke Winkelmann, Kevin Jude, Christoph Thomas, Megan M. Suhoski, Edgar G. Engleman, Jacob Piehler,* K. Christopher Garcia*

*Corresponding author. E-mail: kcgarcia{at}stanford.edu (K.C.G.); piehler{at}uos.de (J.P.)

This PDF file includes:

  • Text
  • Fig. S1. SPR binding sensograms for select IL-13 variants.
  • Fig. S2. Electron density of the site II and site III cytokine binding interfaces in the IL-13 A11–IL-13Rα1–IL-4Rα ternary complex.
  • Fig. S3. Visualization of IL-13–containing ternary complex assembly in live cells.
  • Fig. S4. Co-locomotion trajectories of HaloTag–IL-13Rα1 and SNAPf–IL-4Rα in response to IL-13.
  • Fig. S5. Diffusion properties of HaloTag–IL-13Rα1, SNAPf–IL-4Rα, and the dimerized complex.
  • Fig. S6. STAT6 activation induced by different concentrations of IL-13 variants.
  • Fig. S7. Functional characterization of IL-13 variants.
  • Fig. S8. Analysis of IL-13Rα1 and IL-4Rα abundances in different IL-13–responsive cells.
  • Fig. S9. Endocytosis of IL-13 receptor subunits.
  • Fig. S10. Simulations of STAT6 phosphorylation kinetics and dose-response curves.
  • Table S1. Data collection and refinement statistics (molecular replacement) for the IL-13 A11–IL-13Rα1–IL-4Rα ternary complex.
  • Legends for movies S1 to S6

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Other Supplementary Material for this manuscript includes the following:

  • Movie S1 (.mp4 format). Quantification of receptor density and spatial organization by single-molecule imaging.
  • Movie S2 (.mp4 format). Single-molecule photobleaching experiments.
  • Movie S3 (.mp4 format). Single-molecule imaging of TMRIL-13Rα1 and DY647IL-4Rα.
  • Movie S4 (.mp4 format). Single-molecule imaging of TMRIL-13Rα1 and DY647IL-4Rα.
  • Movie S5 (.avi format). Single-molecule photobleaching of IL-13Rα1 and IL-4Rα.
  • Movie S6 (.mp4 format). Co-locomotion of individual IL-13Rα1–IL-4Rα dimers.

Citation: I. Moraga, D. Richter, S. Wilmes, H. Winkelmann, K. Jude, C. Thomas, M. M. Suhoski, E. G. Engleman, J. Piehler, K. C. Garcia, Instructive roles for cytokinereceptor binding parameters in determining signaling and functional potency. Sci. Signal. 8, ra114 (2015).

© 2015 American Association for the Advancement of Science