CAREER: Dynamics of Computationally Designed Fluorescent Proteins

Wesleyan University, Middletown CT

Investigators

Abstract

This award is funded in whole or in part under the American Rescue Plan Act of 2021 (Public Law 117-2). The design of proteins with novel activity, behavior, or purpose has been an important area of research. The field of computational protein design is becoming increasingly sophisticated, enabling not only the redesign of existing proteins but also the creation of proteins entirely from scratch. The goal of this project is to study and optimize the movement of microscopic, computationally designed proteins that use light to track the locations of biological molecules and reveal the inner working of living organisms. This will enable the creation of even more advanced versions of these and other protein machines that can also help in the manufacture and recycling of materials at the chemical level. The project will employ several graduate and undergraduate students in full-time research, preparing them for careers where computational and experimental methods are increasingly integrated to solve the next set of major scientific challenges. This work will support a suite of new class-based research projects, empowering undergraduates in scientific discovery and advancing knowledge. American Rescue Plan funding is used to support this early career investigator at a critical stage in his career. While the structure of a protein is almost always necessary for function, it is often not sufficient. This project focuses on the critical but often neglected role of protein motion in enabling absorbance and reemission of light, a process known as fluorescence. We will first determine which protein shapes either enhance or inhibit fluorescence through detailed analysis of computer simulations and extensive experimental structural characterization. Second, we will test our models through redesign and experimental examination of brighter fluorescent protein variants. As part of these efforts, we will develop a general-purpose computer algorithm that enables rapid evaluation of how thousands of potential mutations affect the shape of the protein. Third, we will investigate the structural determinants of other important properties like the ability of the protein to prevent or facilitate switching fluorescence on and off. Ultimately, this project aims to develop a detailed understanding of how these fluorescent proteins and others can be redesigned to make them truly useful tools for biological research and industrial processes. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.