2. Academic Validation
  3. AspSnFR: A genetically encoded biosensor for real-time monitoring of aspartate in live cells

AspSnFR: A genetically encoded biosensor for real-time monitoring of aspartate in live cells

  • Cell Chem Biol. 2024 May 20:S2451-9456(24)00179-X. doi: 10.1016/j.chembiol.2024.05.002.
Lars Hellweg 1,  Martin Pfeifer 2,  Miroslaw Tarnawski 3,  Shao Thing-Teoh 4,  Lena Chang 2,  Andrea Bergner 5,  Jana Kress 5,  Julien Hiblot 5,  Tabea Wiedmer 4,  Giulio Superti-Furga 6,  Jürgen Reinhardt 2,  Kai Johnsson 7,  Philipp Leippe 8
Affiliations

Affiliations

  • 1 Department of Chemical Biology, Max Planck Institute for Medical Research, Heidelberg, Germany; Heidelberg University, Heidelberg, Germany.
  • 2 Novartis Biomedical Research, Discovery Science, Basel, Switzerland.
  • 3 Protein Expression and Characterization Facility, Max Planck Institute for Medical Research, Heidelberg, Germany.
  • 4 CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria.
  • 5 Department of Chemical Biology, Max Planck Institute for Medical Research, Heidelberg, Germany.
  • 6 CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria; Center for Physiology and Pharmacology, Medical University of Vienna, Vienna, Austria.
  • 7 Department of Chemical Biology, Max Planck Institute for Medical Research, Heidelberg, Germany; Institute of Chemical Sciences and Engineering (ISIC), École Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland. Electronic address: johnsson@mr.mpg.de.
  • 8 Department of Chemical Biology, Max Planck Institute for Medical Research, Heidelberg, Germany; CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria. Electronic address: pleippe@cemm.oeaw.ac.at.
PMID: 38806058 DOI: 10.1016/j.chembiol.2024.05.002
Abstract

Aspartate is crucial for nucleotide synthesis, ammonia detoxification, and maintaining redox balance via the malate-aspartate-shuttle (MAS). To disentangle these multiple roles of aspartate metabolism, tools are required that measure aspartate concentrations in real time and in live cells. We introduce AspSnFR, a genetically encoded green fluorescent biosensor for intracellular aspartate, engineered through displaying and screening biosensor libraries on mammalian cells. In live cells, AspSnFR is able to precisely and quantitatively measure cytosolic aspartate concentrations and dissect its production from glutamine. Combining high-content imaging of AspSnFR with pharmacological perturbations exposes differences in metabolic vulnerabilities of aspartate levels based on nutrient availability. Further, AspSnFR facilitates tracking of aspartate export from mitochondria through SLC25A12, the MAS' key transporter. We show that SLC25A12 is a rapidly responding and direct route to couple CA2+ signaling with mitochondrial aspartate export. This establishes SLC25A12 as a crucial link between cellular signaling, mitochondrial respiration, and metabolism.

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