Career: Site-Specific Dual-Labeling of A Protein Through Two Genetically Incorporated Noncanonical Amino Acids
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The Chemistry of Life Processes Program funds Professor Wenshe Liu of Texas A & M University for a CAREER award to carry out research entitled "Site-specific dual-labeling of a protein through two genetically incorporated noncanonical amino acids." Studies of protein structural rearrangement and folding/unfolding processes are of great importance for our understanding of protein functions and a lot of human diseases that are caused by the accumulation of misfolded proteins. A general method for protein structural rearrangement and folding/unfolding studies is to analyze fluorescent resonance energy transfer (FRET) between a pair of donor and acceptor dyes that are selectively attached to the protein of interest. However, a major difficulty lies in achieving dually fluorescent labeling of a protein site-specifically. The research goal of this proposal is to develop simple and reliable methods for site-selective dual labeling of a protein with a FRET pair. The proposed methods are based on the genetic incorporation of two different noncanonical amino acids into a protein followed by their selective modifications with a FRET pair afterward. We aim to optimize the dual labeling conditions and develop biosensors for small molecules and enzymes. The intrinsic merit of site-selective dual labeling methods is their simplicity. They require only the conventional and readily accessible recombinant DNA techniques and simple organic synthesis. This feature is expected to promote rapid adoption of the proposed methods by other research groups and accelerate related studies. The education goal of this proposal is to integrate the proposed research into the undergraduate chemistry curriculum by providing training in modern research skills and stimulating critical thinking and creativity. The proposed interdisciplinary research will involve graduate students, undergraduate students and postdoctoral scholars, with particular emphasis on including women and members of underrepresented minorities. A summer research program for undergraduate students from four-year colleges and chemistry lab kits for high school students are expected to help encourage college and high school students to pursue studies and academic careers in STEM areas.
