Developing a Selective Sensor to Detect Carbon Monoxide in Live Cells
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Carbon monoxide has gained considerable interest due to its role as an important signaling molecule in cells. Studying carbon monoxide is difficult, however, due to a lack of reliable carbon monoxide detectors in vivo. CooA is a protein that has the ability to selectively bind to carbon monoxide molecules in the organism R. Rubrum. This protein undergoes a conformational change once bound to carbon monoxide. To develop this sensor, we mutated a codon at a position that likely undergoes a polarity shift once this conformational change takes place. P- azido-phenylalanine is an unnatural amino acid that allows us to perform click chemistry to attach any fluorescent molecule that contains an alkyne group. Alkyne MegaStokes dye 673 is one of these organic molecules that contains an alkyne group, the intensity of fluorescence changes depending on the polarity of the surrounding environment. Thus far, we have mutated CooA in two positions, incorporated the unnatural amino acid into the mutated site(s), and conjugated the Megastokes fluorescent dye to the azide-containing amino acid. The next step is to test the fluorescence change following carbon monoxide binding to the CooA, a change in fluorescence will give us a suitable way to detect carbon monoxide in vivo.
Wetzel, Edward A., "Developing a Selective Sensor to Detect Carbon Monoxide in Live Cells" (2014). Fall Interdisciplinary Research Symposium. Paper 64.
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