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Updated: Jul 19, 2026

Metabolic Labeling of Leucine Rich Repeat Kinases 1 and 2 with Radioactive Phosphate

Metabolic Labeling of Leucine Rich Repeat Kinases 1 and 2 with Radioactive Phosphate

Published on: September 18, 2013

A novel analytical method for in vivo phosphate tracking.

Hong Gu¹, Sylvie Lalonde, Sakiko Okumoto

¹Carnegie Institution, Department of Plant Biology, 260 Panama Street, Stanford, CA 94305, USA.

|October 13, 2006

Summary

New genetically-encoded fluorescence resonance energy transfer (FRET) sensors, called FLIPPi, enable real-time monitoring of inorganic phosphate (P(i)) metabolism in living cells. This tool aids in studying cell transport and metabolic changes.

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Area of Science:

Biochemistry
Molecular Biology
Cell Biology

Background:

Phosphate (P(i)) is crucial for cellular energy and signaling.
Existing methods for measuring intracellular P(i) have limitations.
Genetically-encoded sensors offer advantages for real-time cellular analysis.

Purpose of the Study:

To engineer and validate genetically-encoded fluorescence resonance energy transfer (FRET) sensors for inorganic phosphate (P(i)).
To demonstrate the utility of these sensors for monitoring P(i) dynamics in living cells.

Main Methods:

Engineering of FLIPPi sensors by fusing a phosphate-binding protein (PiBP) to eCFP and Venus fluorophores.
Characterization of sensor FRET efficiency in response to varying P(i) concentrations.

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Published on: January 19, 2017

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Last Updated: Jul 19, 2026

Metabolic Labeling of Leucine Rich Repeat Kinases 1 and 2 with Radioactive Phosphate

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Published on: September 18, 2013

Phosphorus-31 Magnetic Resonance Spectroscopy: A Tool for Measuring In Vivo Mitochondrial Oxidative Phosphorylation Capacity in Human Skeletal Muscle

Phosphorus-31 Magnetic Resonance Spectroscopy: A Tool for Measuring In Vivo Mitochondrial Oxidative Phosphorylation Capacity in Human Skeletal Muscle

Published on: January 19, 2017

Co-expression of FLIPPi sensors with Na(+)/P(i) cotransporters in COS-7 cells to assess P(i) uptake.

Main Results:

FLIPPi sensors exhibited P(i)-dependent increases in FRET efficiency.
Mutant FLIPPi sensors demonstrated a wide dynamic range for P(i) detection (eight orders of magnitude).
Observed FRET changes in cells confirmed concentrative P(i) uptake via the PiT2 transporter.

Conclusions:

Genetically-encoded FLIPPi sensors are effective tools for real-time P(i) monitoring in living cells.
FLIPPi sensors facilitate studies in fluxomics and cellular pathophysiology.
These sensors provide novel insights into P(i) metabolism and transport during cellular processes like migration.