Development of a robust fluorescent ATP sensor and its application for quantitative analysis

2017 Fiscal Year Final Research Report

• Project/Area Number: 16K14024
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Analytical chemistry
• Research Institution: Waseda University
• Principal Investigator: Arai Satoshi 早稲田大学, 総合研究機構, 次席研究員(研究院講師) (70454056)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Keywords: Fluorescent protein / Fluorescent sensor / ATP / Bioimaging / Quantitative analysis / Single cell

Outline of Final Research Achievements

An adenosine triphosphate (ATP) is considered to a most important molecule as a molecular unit of currency in biological studies on energy metabolism. Our study is aimed at establishing a method that allows quantitative analysis of intracellular ATP level at targeted place at single cellular resolution. We generated a genetically encoded fluorescent sensor which comprises a green fluorescent protein variant and the part of the bacterial FoF1-ATP synthase. It exhibited the change in fluorescence as well as fluorescent lifetime in response to the change in ATP concentration. Thanks to this unique property, we were able to obtain the spatio-temporal information regarding ATP dynamics and analyse its concentration quantitatively at the same time at the same cell. More specifically, we succeeded to observe the difference of intra/sub-cellular ATP level between different cell types and organelles.

Free Research Field

Chemical Biology