Hydrazone Reactions as a Tool for Intracellular Bioorthogonal Labeling

Authors: Naomy DeLosSantos, Kamalika Mukherjee

Field of Study: Science, Technology, Engineering, and/or Math

Faculty Mentors: Susan Bane

Easel: 1

Timeslot: Morning

Abstract: Bioorthogonal chemistry has been widely used to explore biological systems through covalent bond formation. Specifically, hydrazone formation has been used to fluorescently label cellular proteins of interest. Previously, this lab has focused on using this chemistry with a hydrazine-functionalized synthetic fluorophore. It exhibits a “turn on” fluorescence, a shift in emission wavelength and an increase in emission intensity upon hydrazone formation. This chemistry was used to site-specifically label proteins of interest on microtubules and for the detection of carbonylation caused by oxidative stress in live cells. In this work, a simple method for detecting protein carbonylation in fixed cells has been developed with a hydrazide-functionalized fluorophore. This fluorophore is commercially available, more chemically stable, and can “turn up” its fluorescence upon hydrazone formation. The utility of the fluorophore in detecting carbonylation was observed in A549 cells, and fluorescent signals were quantified with a fluorescence plate reader.