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Novel High-Dynamic-Range Fluorescent Sensor to Track Intracellular ATP

A new sensor developed at Janelia Research Campus is providing scientists with the best view yet at ATP levels inside living cells. This allows them to study how fluctuations in this cellular currency affect the cell and contribute to disease, according to a study published in Proceedings of the National Academy of Sciences.

The cellular economy depends on ATP. Since almost every cell function depends on the energy-carrying molecule, ATP is essential to cellular life.

Despite ATP’s vital role in cells, scientists have been unable to effectively monitor its alterations in living cells. The ATP sensors of the past were slow, dim, or challenging to operate.

A fluorescent protein sensor called iATPSnFR was created in 2019 by Janelia and UCLA researchers. It functions similarly to the well-known GCaMP sensors used to detect calcium. A fluorescent molecule is tethered to an ATP-binding protein. The protein alters shape as a result of this interaction, which illuminates the fluorescent molecule.

The limited operational range of this first-generation sensor rendered it unsuitable for monitoring changes in ATP concentrations within cells despite its ability to detect ATP changes.

Now, Janelia scientists and collaborators, led by Jonathan Marvin, a senior scientist on Janelia’s Tool Translation Team, have created the next-generation sensor, iATPSnFR2. This sensor can monitor ATP concentrations over a considerably wider range, allowing scientists to detect ATP within living cells with higher precision than ever before.

Tim Ryan, a Weill Cornell Medicine researcher, and Janelia Scholar, who collaborated with Marvin and the team to design and test the novel sensor, utilized iATPSnFR2 to monitor ATP levels at individual synapses, which are the junctions where neurons interact.

Ryan and his team are investigating how changes in ATP activity at synapses can contribute to the development of Parkinson’s disease. The novel sensor allows them to directly see these variations and identify how these fluctuations could be related to the disease.

Beyond this research, the team anticipates that other scientists will utilize the novel sensor to investigate a wide range of ATP-related research problems that have been difficult to address.

Journal Reference:

Marvin, J. S., et al. (2024) iATPSnFR2: A high-dynamic-range fluorescent sensor for monitoring intracellular ATP. Proceedings of the National Academy of Sciences. doi:10.1073/pnas.2314604121.

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