Expanding the capabilities and usage of the TELSAM protein crystallization chaperone

There is a critical need for new protein crystallization methods that are more successful and require less labor, time, and resources. Lack of straightforward methods to successfully crystallize any protein of interest significantly hinders study of molecular disease mechanisms and the development of effective treatments. This lack of effective treatments for many diseases forces them to be addressed instead with costly symptom management programs. Over the past four years, we have investigated TELSAM, a novel polymer-forming protein crystallization chaperone. TELSAM carrier proteins can be genetically fused to disease proteins, drug targets, and bioengineered proteins. In low pH crystallization conditions, TELSAM-target protein fusions polymerize, and the resulting polymers zipper up to form crystals suitable for X-ray diffraction and atomic resolution structure determination. TELSAM fusion readily forms crystals of 90% of proteins of interest (a stark improvement over the 30% crystallization rate of traditional methods) and routinely at protein concentrations of 1 mg/mL (promising to enable the structure determination of proteins that can only be produced in minute quantities). TELSAM fusion crystallography thus has the potential to revolutionize the small molecule and biologic therapeutic industries by accelerating structure determination steps, currently a bottleneck. For TELSAM to realize this potential, academic and industrial structure biologists need 1) experimentally validated guiding principles for the use of TELSAM, 2) a sufficient number of successful use cases to demonstrate general efficacy, and 3) demonstration of the applications and limits of TELSAM fusion crystallization. Thus far we have rigorously investigated the guiding principles for TELSAM’s use and begun to demonstrate its usefulness with proteins relevant in human disease. Our goals for the next five years are to rigorously address the above three needs and broadly disseminate our findings to the structural biology community. The overall vision of our research program is focused on pushing the limits of protein engineering and structure determination fields while at the same time developing undergraduate and graduate students into excellent biochemists. We do this by putting them at the front lines of groundbreaking research. Both undergraduate and graduate students in our group participate in all parts of scientific inquiry, including reagent preparation, experiment design and execution, data collection and analysis, and manuscript preparation and presentation at national and international meetings. The proposed research is significant because it will accelerate the successful structure determination of a greater number and variety of biotechnology and disease-relevant proteins, drugs, and biologics, ultimately leading to new biotechnology tools, more effective disease treatments, and reduced healthcare costs.