Huntley Chang (MASc): Development of genetically encoded sensors based on PKM2

March 14, 2018 @ 9:00 am – 9:30 am
Rosebrugh Building
Rosebrugh Bldg, Toronto, ON M5S 3G9

Room: RS 211


We recently developed a family of genetically encoded sensors based on steady-state fluorescence anisotropy, which we have previously used to track the NADPH/NADP+ redox state of beta-cells in real time (Apollo-NADP+). NADPH is critically used to scavenge toxic reactive oxygen species (ROS), and is also used in the biosynthesis of lipids, amino acids, and nucleotides during proliferation. Normal cell proliferation requires the shutting off of glycolysis to generate metabolic intermediates, which are shunted into the pentose phosphate pathway (PPP). The PPP has an oxidative phase for NADPH generation, and a non-oxidative phase to generate metabolic building blocks. However, due to their lack of significant PPP activity, beta-cells generate most of their NADPH through products of glycolysis instead. Different metabolic states, such as beta-cell proliferation and survival, may influence the activity of these different pathways. A better understanding of metabolic flux is needed to understand how these scenarios could affect beta-cell functionality. We are thus interested in developing another sensor for glycolysis which will enable us to track beta-cell NADPH/NADP+ redox state and glycolysis in tandem.