Supplementary Materials

Supplementary Materials for:

Triple-Color FRET Analysis Reveals Conformational Changes in the WIP-WASp Actin-Regulating Complex

Sophia Fried, Barak Reicher, Maor H. Pauker, Shani Eliyahu, Omri Matalon, Elad Noy, Jordan Chill, Mira Barda-Saad*

*Corresponding author. E-mail: mira.barda-saad@biu.ac.il

This PDF file includes:

  • Fig. S1. Analysis of protein abundance and interactions between wild-type and mutant forms of WIP and WASp and actin.
  • Fig. S2. Phosphorylation of WIP by PKCθ controls WASp ubiquitylation and stability.
  • Fig. S3. The N terminus of WIP and the VCA domain of WASp are required for the WIP-WASp interaction.
  • Fig. S4. Involvement of actin in the WIP-WASp interaction.
  • Fig. S5. Analysis of tagged protein abundance in cells used for 3FRET.
  • Table S1. FRET efficiency and statistical analysis of the interaction of WASp tagged at its N terminus with the N terminus or C terminus of WIP, as well as intramolecular FRET between the two molecular ends of WIP.
  • Table S2. FRET efficiency and statistical analysis of the interaction of WASp tagged at its C terminus with the N terminus or C terminus of WIP, as well as intramolecular FRET between the two molecular ends of WIP.
  • Legends for movies S1 to S5

[Download PDF]

Technical Details

Format: Adobe Acrobat PDF

Size: 839 KB

Other Supplementary Material for this manuscript includes the following:

  • Movie S1 (.mov format). Dynamic movement of CFP-WASp and YFP–WIP wild type in live activated T cells.
  • Movie S2 (.mov format). Dynamic movement of YFP–WIP wild type in live activated T cells.
  • Movie S3 (.mov format). Dynamic movement of YFP–WIP R485,490K in live activated T cells.
  • Movie S4 (.mov format). Dynamic movement of YFP–WIP S488D in live activated T cells.
  • Movie S5 (.mov format). Dynamic movement of CFP-actin and YFP–WIP wild type in live activated T cells.

[Download Movies S1 to S5]


Citation: S. Fried, B. Reicher, M. H. Pauker, S. Eliyahu, O. Matalon, E. Noy, J. Chill, M. Barda-Saad, Triple-Color FRET Analysis Reveals Conformational Changes in the WIPWASp Actin-Regulating Complex. Sci. Signal. 7, ra60 (2014).

© 2014 American Association for the Advancement of Science