Development of genetically encoded heme sensors

Abstract

Due to the biological importance of heme and its implication in various disease states, uncovering how it is transported throughout the cell is of vital importance. Some of the strongest in vivo tools present in the literature are FRET-based sensors using a number of chromophores that are optimized and expanded from GFP. In order to elucidate the movement of heme throughout the cell, GFP FRET -based heme sensors were designed, expressed, and purified to be further characterized in vitro. This series of heme sensors were expressed in Saccharomyces cerevisiae to monitor the in vivo movement of heme. Different growth conditions were explored to monitor the effect of these changes to cytosolic heme availability. These heme sensors are now poised to address the movement of heme from the mitochondria to other targets in the cell under a variety of conditions. This will provide insight into heme trafficking pathways, as well as the role heme plays in neurodegenerative diseases and aging