Aman Popli, “Investigating the Mechanical Behavior of Talin Protein Using Magnetic Tweezers”
Mentor: Ionel Popa, Physics
Poster #147

Single-molecule force spectroscopy with magnetic tweezers is a powerful technique used to examine the behavior of molecules when they are subjected to mechanical force, allowing for analysis of conformational changes, movements, and interactions in biological molecules like proteins and nucleic acids. Force plays a fundamental role in numerous biological processes, and force spectroscopy enables the precise measurement of force and displacement produced by individual molecules, ranging from cells to proteins. The application of mechanical force to a protein can modulate its function and other processes associated with mechanical transduction. In this research, we employ covalent bonding and single-molecule magnetic tweezers to explore the impact of applied mechanical force on the mechanical properties of the talin rod domains R13 and DD. Talin is a cytoskeletal protein, acts as the molecular computer of our cells by mediating interactions between the skeleton of the cell and its extracellular matrix. The protein talin also contributes in cell signaling, cell adhesion and migration. We utilized covalent attachment chemistry employing HaloTag and SpyTag to engineer and express talin rod domains anchored between these two tethering tags. This enabled us to evaluate unfolding and refolding kinetics of R13-DD under forces comparable to those experienced by talin during its function. Under mechanical force, the talin rod domains, R13-DD, attract binding partners like vinculin. The interactions between talin domains and their binding partners are crucial for regulating mechanotransduction processes, which are significant in the development of cancer and metastasis.