Hi all,
I was scanning the literature looking for a method to measure ATP concentrations as a function of time in live bacterial cells (i would like to do that in cyanobacterial organisms). So far I only found the firefly luciferase assay, which involves lysis of the cells. Is possible to apply this assay to live cells maybe expressing the luciferase from a plasmid?are there any different technique for measuring ATP levels in live cells?
Any suggestion or even better literature reference will be most appreciated.
Best,
Jacopo
There was an article on measuring [ATP] in E. coli by expressing a mutant firefly luciferase with a higher Km for ATP, but they have nonetheless permeabilized the cells during the assay:
"Relationship between growth rate and ATP concentration in Escherichia coli: a bioassay for available cellular ATP" (doi: 10.1074/jbc.M311996200)
Also, the following articles touched this topic:
2) "Evidence for rapid consumption of millimolar concentrations of cytoplasmic ATP during rigor-contracture of metabolically compromised single cardiomyocytes" (http://www.biochemj.org/bj/319/0463/bj3190463.htm).
3) "Synchronized ATP oscillations have a critical role in prechondrogenic condensation during chondrogenesis" (doi:10.1038/cddis.2012.20).
So, you can check if a beetle luciferase (some WT or with a high Km for ATP) and luciferin in the medium would work for this.
Hi Jacopo, have you try the QUEEN plasmid. This allows to measure ATP in real time.
https://www.nature.com/articles/srep06522
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