From Yeast to Alzheimer’s: Optimizing Human Pyrin-Only Protein 2 Production in Saccharomyces cerevisiae
Category: Oral Presentation
Author(s): Mia Papantonio, Chloe Alivio, Edward Lue Chee Lip, Kaitlyn Manzanares, Mia Krause, Van Mai, Aryan Advaita, Elena Tran
Presenter(s): Mia Papantonio
Mentors(s): Victor Kasper, Jacob Leavitt, Stephanie Moreira
Alzheimer’s is a prevalent neurodegenerative disease that causes cognitive decline, memory loss, and behavioral changes in individuals typically aged 65 or older. The Nod-Like Receptor Protein 3 (NLRP3) plays a vital role in Alzheimer’s development by inducing an inflammatory response in the brain; human Pyrin-Only Protein 2 (POP2) inhibits the NLRP3 pathway, making it a potentially therapeutic protein. This study aims to assess the expression of human POP2 in model organisms Escherichia coli and Saccharomyces cerevisiae (Baker’s yeast) to maximize protein yield. While E. coli is widely used for recombinant protein expression, eukaryotic S. cerevisiae offers beneficial post-translational modifications, potentially essential for protein folding; human POP2 expression has been successful in E. coli, but has yet to occur in S. cerevisiae. By optimizing expression conditions, this study aims to establish S. cerevisiae as a viable production system. A pre-ordered plasmid containing the human POP2 gene fragment will be used in E. coli expression. Using restriction enzyme digests, the codon-optimized human POP2 gene will be cut from the plasmid and cloned into a yeast expression vector for transformation into strain BY4741 of S. cerevisiae. Ideal growth conditions will be optimized via temperature, incubation time, and media composition. Western blot analysis using a x2 FLAG-tag will allow for protein expression level evaluation in both model organisms. This study lays the groundwork for future neurodegenerative disease research and potential applications in Alzheimer’s treatment by demonstrating the feasibility of expressing human POP2 in S. cerevisiae and providing valuable insight into scalable protein expression strategies.