PALM TCR COMPLEXES · Studying the Structure and Dynamics of TCR nucleated Complexes at the single molecule level
7РП — „Хора“ (Действия „Мария Кюри“)
- Период
- 2012-10-01 → 2016-09-30
- Финансиране от ЕС
- 100 000 €
- Участници
- 1
- Схема
- MC-CIG
Линиите свързват координатора с партньорите.
Накратко на български
Структурата и движението на сигнализиращите комплекси в клетките се анализират чрез свръхразрешаваща микроскопия на единични молекули. Разбирането на тези механизми помага при идентифицирането на начини за намеса при неправилни клетъчни функции и заболявания.
Кратко обяснение, генерирано от езиков модел по текста на CORDIS. Оригиналът е по-долу.
Резултати накратко
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. Homepage: http://www.phys.huji.ac.il/ShermanLab/
Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз
Цел на проекта
The cell transduces information across its plasma- membrane via the engagement of surface receptors and the subsequent recruitment of intracellular proteins to these receptors or to adapter proteins to form transient and heterogeneous signaling complexes. Multi- molecular signaling complexes can, in turn, be found in large microclusters or aggregates. Such signaling complexes and clusters have been shown to play a crucial role in T cell activation and thus, in the ability of the immune system to adequately respond to foreign pathogens. However, little is known about the detailed structure, content, and organization of signaling complexes due to severe limitations of current experimental techniques. I propose to develop and apply cutting-edge super-resolution microscopy techniques, biophysical models, and statistical methods to study mechanisms of cell activation by signaling complexes in single-molecule detail in intact cells. Previously, by imaging complexes downstream of the T cell receptor at the single- molecule level with photoactivated localization microscopy (PALM), I found that these complexes show functionally significant nano-scale organization at the plasma- membrane of activated cells. I hypothesize that signaling complexes have additional levels of dynamic organization that are crucial to the plurality of functions of these complexes in adequately activating T cells. Importantly, my developed techniques and expected results will be relevant to many other signaling systems and will greatly extend our understanding of the composition, structure, and formation of signaling complexes and mechanisms of cell activation in health and disease. Such knowledge will present novel opportunities for pharmacological intervention in diseases involving inadequate immune responses and in cancer.
Оригинален текст от CORDIS (на английски).
Участници
- THE HEBREW UNIVERSITY OF JERUSALEM · JerusalemКоординаторИзраел
Връзки
Данни: CORDIS, © Европейски съюз