A novel analytical method for in vivo phosphate tracking
Research output: Contribution to journal › Journal article › Research › peer-review
Genetically-encoded fluorescence resonance energy transfer (FRET) sensors for phosphate (Pi) (FLIPPi) were engineered by fusing a predicted Synechococcus phosphate-binding protein (PiBP) to eCFP and Venus. Purified fluorescent indicator protein for inorganic phosphate (FLIPPi), in which the fluorophores are attached to the same PiBP lobe, shows Pi-dependent increases in FRET efficiency. FLIPPi affinity mutants cover Pi changes over eight orders of magnitude. COS-7 cells co-expressing a low-affinity FLIPPi and a Na+/Pi co-transporter exhibited FRET changes when perfused with 100µM Pi, demonstrating concentrative Pi uptake by PiT2. FLIPPi sensors are suitable for real-time monitoring of Pi metabolism in living cells, providing a new tool for fluxomics, analysis of pathophysiology or changes of Pi during cell migration.
Original language | English |
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Journal | FEBS Letters |
Volume | 580 |
Issue number | 25 |
Pages (from-to) | 5885-5893 |
Number of pages | 9 |
ISSN | 0014-5793 |
DOIs | |
Publication status | Published - 2006 |
Bibliographical note
FLIPPi, fluorescent indicator protein for inorganic phosphate, FRET, fluorescence resonance energy transfer, FP, fluorescent protein
- Former LIFE faculty - Fluorescence energy transfer, Phosphate starvation, Biosensor, Synechococcus
Research areas
ID: 8042551