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FEBS Lett. 2006 Oct 30;580(25):5885-93. Epub 2006 Oct 2.

A novel analytical method for in vivo phosphate tracking.

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

Erratum in

  • FEBS Lett. 2007 Feb 6;581(3):579.

Abstract

Genetically-encoded fluorescence resonance energy transfer (FRET) sensors for phosphate (P(i)) (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 P(i)-dependent increases in FRET efficiency. FLIPPi affinity mutants cover P(i) changes over eight orders of magnitude. COS-7 cells co-expressing a low-affinity FLIPPi and a Na(+)/P(i) co-transporter exhibited FRET changes when perfused with 100 microM P(i), demonstrating concentrative P(i) uptake by PiT2. FLIPPi sensors are suitable for real-time monitoring of P(i) metabolism in living cells, providing a new tool for fluxomics, analysis of pathophysiology or changes of P(i) during cell migration.

PMID:
17034793
[PubMed - indexed for MEDLINE]
PMCID:
PMC2748124
Free PMC Article
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