Approaches to monitor ATP levels in living cells: where do we stand?

Abstract

ATP is the most universal and essential energy molecule in cells. This is due to its ability to store cellular energy in form of high energy phosphate bonds, which are extremely stable and readily usable by the cell. This energy is key for a variety of biological functions such as cell growth and division, metabolism, signaling, and for the turnover of biomolecules. Un-ACCEPTED MANUSCRIPT -CLEAN COPY 2 derstanding how ATP is produced and hydrolyzed with a spatiotemporal resolution is necessary to understand its functions both in physiological and pathological contexts. In this review, we will first describe the organization of the electron transport chain and ATP synthase, the main molecular motor for ATP production in mitochondria. Second, we will review the biochemical assays currently available to estimate ATP quantities in cells, and we will compare their readouts, strengths and weaknesses. Finally, we will explore the palette of geneticallyencoded biosensors designed for microscopy-based approaches, and show how their spatiotemporal resolution opened up the possibility to follow ATP levels in living cells.

Genetically Encoded ATP Biosensors for Direct Monitoring of Cellular ATP Dynamics

“…Upon ATP binding, the two α-helices interact and refine their conformational structure of the ϵ subunit, leading to fluorescent/bioluminescent illumination, indicating that ATP is present. Overall, this subunit adopts two different conformations: open (ATP-free) or closed (ATP-bound) [ 142 , 143 , 144 ]. The uses and applicability of this technology are limitless.…”

Biomaterial-assisted targeted and controlled delivery of CRISPR/Cas9 for precise gene editing

“…The ATP amount is 1000 times more in the intracellular environment than in the outer cell environment. 56 The concentration of the ATP is a better option for developing the CRISPR/Cas9 for targeting low- and high-ATP cells in living organisms. Zinc nanoparticles, ZIF-90, were designed for targeting the ATP-rich cells.…”

Red blood cells in type 1 diabetes and multiple sclerosis and technologies to measure their emerging roles

“…First, both ATP and NO are highly reactive molecules. ATP, with its multiple phosphate groups, can undergo hydrolysis to form ADP or AMP [ 152 ]. NO is a relatively unstable molecule.…”

“…In the 1960s, the luciferin/luciferase assay began being used to quantify ATP in RBCs [ 161 ]. In this commonly used assay, the luciferase enzyme catalyzes a bioluminescent reaction with its substrate luciferin and ATP to produce oxyluciferin at an excited state [ 152 ]. As oxyluciferin relaxes to its ground state, yellow-green light is produced with a maximum emission between 550 and 570 nm, which can easily be detected with a photomultiplier tube (PMT).…”