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Sci. Signal., 28 September 2010
[DOI: 10.1126/scisignal.2000980]

Supplementary Materials for:

Coupling Mechanism of a GPCR and a Heterotrimeric G Protein During Chemoattractant Gradient Sensing in Dictyostelium

Xuehua Xu, Tobias Meckel, Joseph A. Brzostowski, Jianshe Yan, Martin Meier-Schellersheim, Tian Jin*

*To whom correspondence should be addressed. E-mail: tjin{at}niaid.nih.gov

This PDF file includes:

  • Methods
  • Technical details for simulations
  • Fig. S1. Imaging of single molecules of YFP with TIRF microscopy.
  • Fig. S2. Track lengths obtained from tracking cAR1-YFP and Gβ-YFP by SPT.
  • Fig. S3. Individual steps in the analysis of the mobilities of single molecules.
  • Fig. S4. cAR1-YFP proteins remain on the membrane after stimulation with cAMP.
  • Fig. S5. Effects of GTPase activities on the activation kinetics of G proteins.
  • Fig. S6. Simulated adaptation of the PIP3 response of cells exposed to a homogeneous, constant concentration of cAMP.
  • Fig. S7. Simulated PIP3 responses of cells exposed to a rapidly induced, but then constant, extracellular 2:1 gradient of cAMP.
  • Fig. S8. Diffusion constants and region size of cAR1-YFP and Gβ-YFP at the front and back of a cell exposed to a gradient of cAMP.
  • Description of Movies S1 to S4
  • GproteinCoupling.zip file description
  • References

[Download PDF]

Technical Details

Format: Adobe Acrobat PDF

Size: 1,636 KB

Other Supplementary Material for this manuscript includes the following:

  • GproteinCoupling.zip file. This file contains the G protein coupling model in the form of a directory that can be read by the Simmune software. To use this file, unzip it and open the directory with the Simmune modeler. The modeler and simulator can be obtained from http://www3.niaid.nih.gov/labs/aboutlabs/psiim/computationalBiology/.
  • Movie S1 (.avi format in zipped archive). Imaging of single YFP molecules by a TIRF microscope at 30 frames per second.
  • Movie S2 (.avi format in zipped archive). Imaging of cAR1-YFPs on the membrane of a live cell at 30 frames per second.
  • Movie S3 (.avi format in zipped archive). Imaging of Gβ-YFPs on the membrane of a live cell at 30 frames per second.
  • Movie S4 (.avi format in zipped archive). Tracking of spots of single molecules to obtain their position (x, y) and intensity in the movies.
Citation: X. Xu, T. Meckel, J. A. Brzostowski, J. Yan, M. Meier-Schellersheim, T. Jin, Coupling Mechanism of a GPCR and a Heterotrimeric G Protein During Chemoattractant Gradient Sensing in Dictyostelium. Sci. Signal. 3, ra71 (2010).

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