Abstract by Yonghoo Jo
Chemistry and Biochemistry
Optimization of the fluorescent ATP/ADP ratio sensor PercevalHR in vivo in Trypanosoma brucei
Trypanosoma brucei is a kinetoplast parasite transmitted by tsetse fly bites that causes a neurodegenerative disease known as African Sleeping Sickness with limited treatment options. Since glycolysis is the sole energy source of the bloodstream form (BSF) of the parasite, our lab is investigating kinetoplast metabolism using T. brucei as a model. This is done using genetically encoded biosensors which are sensitive to changes in the concentration of a target ligand. This allows us to measure in vivo changes to metabolite concentrations in the cytosol of T. brucei through quantitative measurements via flow cytometry. One of these sensors, PercevalHR, is an ATP/ADP ratio sensor that changes fluorescent emission ratio between violet and blue excitation in response to changes in levels of the two nucleotides. By measuring this, our lab will be able to use fluorescence measurements to determine if the sensor accurately reports decreases in relative ATP levels in vivo from known starvation conditions such as glucose depletion or 2-DOG poisoning.