2025 AIChE Annual Meeting

(183t) Engineered Proteoliposomes for the Elucidation of Protein Activity

Authors

Marjon Zamani - Presenter, Massachusetts Institute of Technology
Ariel Furst, Massachusetts Institute of Technology
Membrane proteins are ubiquitous and necessary to sustain life. When they malfunction, they lead to diseases that are challenging to treat due to the lack of understanding of their underlying failure mechanisms. Currently, in vitro models of membrane proteins fail to recapitulate native protein function, largely due to the interplay of innumerable, cofounding variables in vivo. To enhance the accuracy of in vitro models, we have created a reductionist approach to study membrane protein activity in physiologically relevant environments. Our approach entails engineering proteoliposomes via lipid-enhanced, cell- free protein expression. As an initial demonstration, we reconstitute drug-metabolizing enzymes and demonstrate their native enzymatic metabolism of xenobiotics for the first time. This is a platform technology that can be used to reconstitute a wide range of membrane proteins for applications ranging from drug discovery to disease modeling.