Hellweg, Lars, Pfeifer, Martin, Chang, Lena, Tarnawski, Miroslaw, Bergner, Andrea, Kress, Jana, Hiblot, Julien, Reinhardt, Juergen, Johnsson, Kai and Leippe, Philipp (2024) AspSnFR: A genetically encoded biosensor for real-time monitoring of aspartate in live cells. Cell chemical biology, 31. pp. 1-13. ISSN 2451-9456

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 withmitochondrial aspartate export. This establishes
SLC25A12 as a crucial link between cellular signaling, mitochondrial respiration, and metabolism.

Item Type:	Article
Keywords:	iAspSnFR, SLC Transporter, ReSolute IMI
Date Deposited:	08 Jun 2024 00:45
Last Modified:	08 Jun 2024 00:45
URI:	https://oak.novartis.com/id/eprint/50291