An Improved ATP FRET Sensor for Yeast Shows Heterogeneity during Nutrient Transitions

Dennis Botman, Johan H. Van Heerden, Bas Teusink*

*Corresponding author for this work

Research output: Contribution to JournalArticleAcademicpeer-review


Adenosine 5-triphosphate (ATP) is the main free energy carrier in metabolism. In budding yeast, shifts to glucose-rich conditions cause dynamic changes in ATP levels, but it is unclear how heterogeneous these dynamics are at a single-cell level. Furthermore, pH also changes and affects readout of fluorescence-based biosensors for single-cell measurements. To measure ATP changes reliably in single yeast cells, we developed yAT1.03, an adapted version of the AT1.03 ATP biosensor, that is pH-insensitive. We show that pregrowth conditions largely affect ATP dynamics during transitions. Moreover, single-cell analyses showed a large variety in ATP responses, which implies large differences of glycolytic startup between individual cells. We found three clusters of dynamic responses, and we show that a small subpopulation of wild-type cells reached an imbalanced state during glycolytic startup, characterized by low ATP levels. These results confirm the need for new tools to study dynamic responses of individual cells in dynamic environments.

Original languageEnglish
Pages (from-to)814-822
Number of pages9
JournalACS Sensors
Issue number3
Early online date20 Feb 2020
Publication statusPublished - 27 Mar 2020


  • ATP
  • biosensor
  • budding yeast
  • FRET
  • heterogeneity
  • single-cell


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