Project Description
The main objective of this project is to use a new fluorescence-based approach developed by one of my graduate students to investigate the mechanical unfolding of proteins inside hydrogels. In this project, low volumes of proteins (~5 uL) are transformed into hydrogels of cylindrical shape using Teflon tubes as the templates (diameter ~0.28 mm). A hydrogel is tethered between two hooks, connected to a voice-coil motor and a force sensor. An analog proportional-integral-differential (PID) system compares the force measured by the force sensor, with the setpoint sent from the computer, and adjusts the gel extension by moving the voice coil to minimize the difference between the two inputs. This ‘clamping’ of the force now allows for new types of experiments for measuring the biomechanics of proteins. We have recently upgraded our force-clamp rheometer to operate on-top of an inverted microscope. An EMCCD camera measure the fluorecence signal coming from the fluid chamber. To quantify domain unfolding using fluorescence, we will implement a new method based on 1-anilinonaphthalene-8-sulphonate (ANS), a hydrophobic ‘dye’ which binds to the exposed secondary structure elements of proteins.
Tasks and Responsibilites
1. Fabricate protein hydrogels and prepare needed buffer solution
2. Perform experiments using these fabricated hydrogels and help develop the new setup
3. Maintain consistent communication with the faculty mentor and advisors
4. Model appropriate academic attitudes and behaviors to staff and faculty
5. Attends mandatory weekly group meetings
6. Prepare poster presentations/talks, as required by your funding program.