A highly responsive pyruvate sensor reveals pathway-regulatory role of the mitochondrial pyruvate carrier MPC.

ELife
R Arce-MolinaAlejandro San Martín

Abstract

Mitochondria generate ATP and building blocks for cell growth and regeneration, using pyruvate as the main substrate. Here we introduce PyronicSF, a user-friendly GFP-based sensor of improved dynamic range that enables real-time subcellular quantitation of mitochondrial pyruvate transport, concentration and flux. We report that cultured mouse astrocytes maintain mitochondrial pyruvate in the low micromolar range, below cytosolic pyruvate, which means that the mitochondrial pyruvate carrier MPC is poised to exert ultrasensitive control on the balance between respiration and anaplerosis/gluconeogenesis. The functionality of the sensor in living tissue is demonstrated in the brain of Drosophila melanogaster larvae. Mitochondrial subpopulations are known to coexist within a given cell, which differ in their morphology, mobility, membrane potential, and vicinity to other organelles. The present tool can be used to investigate how mitochondrial diversity relates to metabolism, to study the role of MPC in disease, and to screen for small-molecule MPC modulators.

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Citations

Nov 17, 2020·Frontiers in Cellular Neuroscience·Susanne KöhlerUlrike Winkler
Aug 11, 2020·Seminars in Cell & Developmental Biology·James Ak LeeScott P Allen
Sep 16, 2020·Current Opinion in Biotechnology·César Carrasco-LópezJosé L Avalos
Feb 10, 2021·Nature Chemical Biology·Yusuke NasuRobert E Campbell
Feb 8, 2021·Trends in Cell Biology·Aastha Garde, David R Sherwood
Mar 3, 2021·Cell Metabolism·Nicholas J RossiterJason R Cantor
Aug 26, 2021·ACS Chemical Biology·John N KobersteinMichael S Cohen
Aug 28, 2021·Frontiers in Pharmacology·Vojtech DvorakGiulio Superti-Furga

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Methods Mentioned

BETA
FRET
transfection
fluorescence microscopy
PCR

Software Mentioned

ImageJ
PyronicSF
ARRIVE
SigmaPlot
Berkeley Madonna
M4T
Process Image Calculator / Add
Pyronic

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