RUI: Structural and Functional Investigations of Ferritin Heteropolymers Using a Novel Expression System

Suny College At Potsdam, Potsdam NY

Investigators

Abstract

Most organisms have evolved mechanisms to maintain a balance between iron as an essential nutrient and iron as a potential toxin. Ferritins are multi-subunit iron storage and detoxification proteins that play a central role in the biological management of iron. The goal of this project is to express and purify recombinant isoferritins using a novel expression system, and for the first time, correlate structure-function relationships between recombinant isoferritins and native isoferritins extracted from animal tissues and organs. Isoferritins are complex molecules with exquisite architectural nanostructures made up of two distinct subunit types, named H and L. Despite their discovery more than 8 decades ago, no studies have ever been performed on recombinantly produced isoferritins, nor has any research group been able to synthesize ferritins that mimic native isoferritins in-vivo. Understanding the functional differences between various isoferritin populations, and how different proportions of H- and L-subunits affect the biochemical and functional properties of isoferritins is of critical importance to understanding the biological processing of iron in mammals. This work will also provide undergraduate students training in modern biochemical and biophysical techniques, and laboratory skills necessary to address challenging questions about iron homeostasis, and preparation for future STEM careers in academia or industries. Throughout this project, undergraduate students will be involved in original research pertaining to the biosynthesis, purification, and characterizing of isoferritins of different subunits composition, using a novel expression system that provides any desired combinations of H- and L-subunits within the ferritin molecule, mimicking native isoferritins found in various tissues and organs. To achieve these goals, a combination of molecular biology techniques, light absorbance and fluorescence spectroscopy, capillary electrophoresis, differential scanning calorimetry, Mossbauer spectroscopy, magnetometry, and high-resolution scanning and transmission electron microscopy will be employed. The extensive investigation outlined in this project should provide fundamental insights into isoferritins structure-function relationships and the dynamics of iron metabolism, both at the systemic and the cellular levels. 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.