Infrared Laser Heating Reveals a Reversible Phase Transition of Phosphofructokinase 1 in Mammalian Cells
- 1Lehrstuhl für Biophysikalische Chemie and Research Center Chemical Sciences and Sustainability Research Alliance Ruhr Bochum Germany
Abstract
Metabolic enzymes can operate in concerted networks, so‐called metabolons. The glucosome is a multi‐enzyme complex that includes enzymes of glycolysis and gluconeogenesis and regulates glucose metabolism in mammalian cells. The mechanism of how glucosomes assemble remains largely unknown. Fluorescence recovery after photobleaching (FRAP) experiments suggest that glucosomes are liquid‐like, but direct evidence of a phase transition mechanism is missing. Here, we developed a novel method to measure the phase transition of condensates in living cells by combining infrared laser heating with confocal microscopy of fluorescently labeled phosphofructokinase 1 liver‐type (PFKL). We showed that glucosomes form and dissolve reversibly in HeLa cells upon temperature modulation, allowing us to determine the condensation temperature. We studied cells under osmotic stress to show that glucosomes retain their phase behavior despite a significant decrease in the mobility of PFKL. We propose that this method is a versatile tool to study the phase behavior of biomolecular condensates, yielding complementary information to well‐established techniques such as FRAP.
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BibTeX
@article{mosbach2026infrared,
title = {{Infrared Laser Heating Reveals a Reversible Phase Transition of Phosphofructokinase 1 in Mammalian Cells}},
author = {Morten Mosbach and Sneha Pavanan Kuruvanthara and Lukas Johannknecht and Simon Ebbinghaus},
journal = {Chemistry (Weinheim an Der Bergstrasse, Germany)},
year = {2026},
month = aug,
doi = {10.1002/chem.71500},
url = {https://doi.org/10.1002/chem.71500},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Infrared Laser Heating Reveals a Reversible Phase Transition of Phosphofructokinase 1 in Mammalian Cells
AU - Morten Mosbach
AU - Sneha Pavanan Kuruvanthara
AU - Lukas Johannknecht
AU - Simon Ebbinghaus
PY - 2026
DA - 2026/08/03
JO - Chemistry (Weinheim an Der Bergstrasse, Germany)
DO - 10.1002/chem.71500
UR - https://doi.org/10.1002/chem.71500
AB - Metabolic enzymes can operate in concerted networks, so‐called metabolons. The glucosome is a multi‐enzyme complex that includes enzymes of glycolysis and gluconeogenesis and regulates glucose metabolism in mammalian cells. The mechanism of how glucosomes assemble remains largely unknown. Fluorescence recovery after photobleaching (FRAP) experiments suggest that glucosomes are liquid‐like, but direct evidence of a phase transition mechanism is missing. Here, we developed a novel method to measure the phase transition of condensates in living cells by combining infrared laser heating with confocal microscopy of fluorescently labeled phosphofructokinase 1 liver‐type (PFKL). We showed that glucosomes form and dissolve reversibly in HeLa cells upon temperature modulation, allowing us to determine the condensation temperature. We studied cells under osmotic stress to show that glucosomes retain their phase behavior despite a significant decrease in the mobility of PFKL. We propose that this method is a versatile tool to study the phase behavior of biomolecular condensates, yielding complementary information to well‐established techniques such as FRAP.
SN - 0947-6539
SN - 1521-3765
ER -