Reactive oxygen species (ROS) play a vital role in normal physiological processes as well as in pathological conditions. Reagents that can generate tunable, well-controlled intracellular concentrations of ROS are a critical tool to quantify effects of these species on cellular function. To this end, we have engineered a set of oxygen-activating enzymes. In vitro activity assays have revealed that mutants in this set produce a variety of ROS product ratios and are characterized by TTNs that vary from mutant to mutant. The focus of this poster presentation is the observation and quantification of intracellular enzyme activities in mammalian cell lines using both small molecule and genetically-encoded ratiometric fluorescent sensors.
This preliminary program is subject to change.
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Intracellular Reactive Oxygen Species (ROS) Generation & Quantification Using Engineered Redox Enzymes
Reactive oxygen species (ROS) play a vital role in normal physiological processes as well as in pathological conditions. Reagents that can generate tunable, well-controlled intracellular concentrations of ROS are a critical tool to quantify effects of these species on cellular function. To this end, we have engineered a set of oxygen-activating enzymes. In vitro activity assays have revealed that mutants in this set produce a variety of ROS product ratios and are characterized by TTNs that vary from mutant to mutant. The focus of this poster presentation is the observation and quantification of intracellular enzyme activities in mammalian cell lines using both small molecule and genetically-encoded ratiometric fluorescent sensors.