Protein biogenesis involves the progression of a polypeptide chain through a series of folding intermediates in search for the most thermodynamically stable conformation - the native 3D structure. Taking place in the crowded cellular milieu, this process can lead to native folding or off-pathway misfolded species. Protein misfolding can result in destructive consequences for the cell as it not only impairs functions but can also favor formation of aggregates. A folding quality control system balances synthesis, folding and degradation of proteins thereby maintaining protein homeostasis in eukaryotic cell. Manipulating protein expression to increase protein production perturbs protein homeostasis and results in accumulation of misfolded proteins and formation of aggregates. Misfolding and aggregation, in turn, cause induction of proteotoxic stress and activation of apoptosis. The accumulation of misfolded proteins favor misfolding of newly synthesized proteins, further compromising folding and stability of the proteome, and the formation of aggregates is highly cytotoxic. Not surprisingly, protein misfolding and aggregation are key impediments to the high-yield production of recombinant proteins, which is one of the fastest growing sectors of the pharmaceutical industry.
The molecular mechanisms that regulate the disposal of misfolded proteins still remain elusive. This project will address these fundamental cell biology questions and will make use of synthetic biology approaches and protein engineering techniques to decipher and manipulate the innate cellular degradation capacity. Results from this work will enable the design of novel cell engineering strategies to limit accumulation of misfolded proteins, prevent aggregation and enhance production of natively folded proteins. These strategies have the potential to significantly impact the high yield production of recombinant proteins for research, industrial and diagnostic applications. In addition, the knowledge gained from this study will advance our understanding of aging and protein misfolding diseases.
The proposed experimental plan will provide a setting to educate undergraduate students. It will also create an effective platform for reaching out to high school students and inspire their interest in science and technology with the goal to increase and diversify the pool of students interested in biotechnology related disciplines. The educational plans incudes creating a series of mentoring programs and research experiences to engage undergraduate and high school students, with particular emphasis to women and underrepresented minorities.
