Supplementary Materials

Characterization of the Intrinsic and TSC2-GAP�Regulated GTPase Activity of Rheb by Real-Time NMR

Christopher B. Marshall, Jason Ho,1 Claudia Buerger, Michael J. Plevin, Guang-Yao Li, Zhihong Li, Mitsuhiko Ikura,* Vuk Stambolic*

This PDF file includes:

  • Methods
  • Fig. S1. Role of Rheb in mTOR signaling.
  • Fig. S2. Spectra of Rheb-GDP and Rheb-GMPPNP.
  • Fig. S3. Secondary structure and chemical shift index analysis.
  • Fig. S4. Preparation of Rheb-GTP for GTPase assay.
  • Fig. S5. Time course of GTP hydrolysis of Rheb-GTP.
  • Fig. S6. Analysis of Rheb-GTP and Rheb-GDP during GTP hydrolysis.
  • Fig. S7. Circular dichroism spectra of WT TSC21225�1742 and its variants.
  • Fig. S8. Further development of real-time NMR method to increase temporal
  • resolution.
  • Fig. S9. Further development of real-time NMR method to decrease sample requirement.
  • References

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1Division of Signaling Biology, Ontario Cancer Institute, University Health Network, Toronto, Ontario, Canada M5G 2M9. 2Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada M5G 2M9.

*To whom correspondence should be addressed. E-mail, vuks{at}uhnres.utoronto.ca (V.S.) and mikura{at}uhnresearch.ca (M.I.)

Citation: C. B. Marshall, J. Ho, C. Buerger, M. J. Plevin, G.-Y. Li, Z. Li, M. Ikura, V. Stambolic, Characterization of the Intrinsic and TSC2-GAP�Regulated GTPase Activity of Rheb by Real-Time NMR. Sci. Signal.2, ra3 (2009).

© 2009 American Association for the Advancement of Science