Excellence in Research: Development of a Fluorescent Assay to Measure LAT1 Transport

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With the support of the Chemistry of Life Processes Program in the Chemistry Division and the Historically Black Colleges and Universities Excellence in Research (HBCU-EiR) Program, Dr. Jacqueline Smith will investigate the transport of small molecules through native transporters. Cell survival depends on the delivery of nutrients such as amino acids, carbohydrates and other valuable biochemicals into the cell. The delivery of the nutrients is enabled by transporters, which are proteins located in the cell membrane. The study of these proteins is challenging. Dr. Smith’s research seeks to create novel chemical probes to study a transporter, specifically the large amino acid transporter (LAT1). The research will also focus on the development of a method for rapid identification of small molecules that are substrates for LAT1. This project will give students at Bowie State an opportunity to explore protein biochemistry and other modern chemical biology techniques. The large amino acid transporter (LAT1) is a key nutrient transporter that is highly expressed in the cells in the brain and placenta and in cells with high energy demand in general. As a transmembrane protein, the structure of LAT1 has been determined only recently and transport by LAT1 was traditionally measured using radioisotope assays. Although computational methods have been used to study LAT1, experimental methods for validation of the computational predictions are lacking. Dr. Smith will pursue the development of proteoliposomes that contain LAT1. The small molecule binding to LAT1 within the proteoliposomes will be studied by surface plasmon resonance (SPR). Newly developed fluorescent substrates will be used to screen for molecules that can be transported through LAT1. The tools developed in this study will be applied to gain a better understanding of the binding to and transport of small molecules by LAT1.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.