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Studying the Structure and Dynamics of TCR nucleated Complexes at the single molecule level

Final Report Summary - PALM TCR COMPLEXES (Studying the Structure and Dynamics of TCR nucleated Complexes at the single molecule level)

In the laboratory for Biophysics at the Racah Institute of Physics, we are interested in the combination of technique development, experiments, computational modeling and simulations to resolve critical mechanisms of cell activation and function in single molecule detail. Ongoing efforts in our lab include:

* Developing techniques for multi-color, Live-cell single-molecule super resolution microscopy
In our lab, we develop and apply combinations of single molecule localization microscopy (SMLM), including photoactivated localization microscopy (PALM) and direct stochastic optical reconstruction (dSTORM) in multiple colors. These methods enable the study of signaling complexes in single molecule detail in intact (fixed and live) cells with resolution down to ~20nm. We further combine and synergize these techniques with related techniques, such as Super-resolution Optical Fluctuation Imaging (SOFI) and single particle tracking (SPT), for optimal live cell imaging and physical characterization at the single molecule level.

*Resolving the cooperativity and structure of signaling complexes
Single molecule super-resolution imaging of three molecular species in locations of high molecular densities has allowed us to study the complexity of molecular interactions, including potential cooperativity or competition in molecular binding.

* Studying biophysical mechanisms of cell activation at the single molecule level
We aim to develop a fundamental understanding of critical mechanisms of cell activation in health and disease in single molecule detail. Ultimately, such a level of understanding could serve to identify novel and efficient ways of intervening in aberrant signaling pathways and cellular malfunctions. To overcome current limitations in research techniques, we rely on cutting-edge microscopy techniques at the single molecule level of intact cells on functionalized interfaces, advanced statistical methods and quantifiable models based on physics of complex systems. Research in our lab is currently focused on the activation of T cells, which plays a central role in mounting adequate immune responses to foreign pathogens. Previous work has resulted in several intriguing findings, such as that signaling complexes at the plasma membrane of T cells have nanoscale structure and organization that facilitate intact cell activation.

Selected Publications from our lab:

V. A. Barr, E. Sherman, J. Yi, I. Akpan, A. K. Rouquette-Jazdanian and L. E. Samelson Development of nanoscale structure in LAT-based signaling complexes, J Cell Sci, Accepted for publication (2016).
Sherman, E.1 Barr, V., Merrill, R. K., Regan, C. K., Sommers, C. L., Samelson, L. E. (2016). Hierarchical nano-structure and synergy of multi-molecular signaling complexes. Nature Communications, 7:12161. 1 - Corresponding author

Sherman, E. (2016). Resolving protein interactions and organization downstream the T cell antigen receptor using single-molecule localization microscopy: a review. Methods Appl. Fluoresc. 4 022002.
Schidorsky, S.*, Yi, X.*, Razvag, Y., Golan, Weiss, S.1 Sherman, E.1Synergizing superresolution optical fluctuation imaging with single molecule localization microscopy. arXiv: 1603.04028. * - Equal contribution. 1 - Corresponding authors

Dubey, G. P.*, Mohan, G. B. M.*, Dubrovsky, A., Amen, T., Tsipshtein, S., Rouvinski, A., Rosenberg, A., Kaganovich, D., Sherman, E., Medalia, O., and Ben-Yehuda, S. (2016). Architecture and Characteristics of Bacterial Nanotubes. Developmental Cell, 36 (4) 452-61. * - Equal contribution.

Neve-Oz, Y. Razvag, J. Sajman, and E. Sherman. Mechanisms of localized activation of the T cell antigen receptor inside clusters. Biochim. Biophys. Acta. 1853 (2015), p. 810–21.

Parker J., Sherman E., van de Raa M., van der Meer D., Samelson L. E., & Losert W. (2013). Automatic sorting of point pattern sets using Minkowski Functionals. Phys. Rev. E., 88, 022720.

Sherman, E., Barr, V., Samelson, L. E. (2013). Resolving multi-molecular protein interactions by photoactivated localization microscopy. Methods, 59 (3), 261-9.