EAGER: Developing Optically Triggered Protein Actuators in Living Organisms

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With this award, the Chemistry of Life Processes Program of the Chemistry Division and the Cellular Dynamics and Function Cluster of the Division of Molecular and Cellular Biosciences are funding Prof. Khalid Salaita from Emory University to develop and characterize a new class of genetically-encoded proteins that exert a mechanical force when optically excited. Despite the importance of mechanotransduction, the molecular details relating the magnitude, timing, and location of forces to specific biochemical pathways remain poorly understood. This is due, in part, to the inability to control mechanical tension within individual proteins. Therefore the precise role of mechanics in regulating biochemical pathways remains difficult to test. The research program led by Prof. Salaita aims to address this limitation and to aid in the integration of the growing fields of optogenetics, mechanobiology, and biophysics. To maximize the dissemination of the research results and to increase adoption of the new tools by researchers in the area of mechanobiology, the methods describing the optimization and biophysical response of force actuator module variants are made available through publication in scientific journals and will be also made public in a session dedicated to opto-mechanics at an upcoming regional molecular and cellular biophysics meeting. The project provides highly multidisciplinary, hands-on training for graduate and undergraduate students.The proposed research focuses on the development of a new class of genetically-encoded proteins that exert a mechanical force when illuminated. Upon illumination, these "force actuators" are expected to collapse and apply a tension of specific magnitude with high spatial and temporal resolution and in a site-selective and non-invasive manner. The opto-mechanical properties of the fluorescently-tagged force actuators are characterized by steady-state and time-resolved spectroscopic methods. For initial in-vitro studies, the force actuator modules are introduced between the head and tailpiece of vinculin. Vinculin was chosen because it is a core protein within focal adhesion complexes and the tension across vinculin regulates a range of processes such as cell protrusion and migration.