| Literature DB >> 17034793 |
Hong Gu1, Sylvie Lalonde, Sakiko Okumoto, Loren L Looger, Anne Marie Scharff-Poulsen, Arthur R Grossman, Jens Kossmann, Iver Jakobsen, Wolf B Frommer.
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.Entities:
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Year: 2006 PMID： 17034793 PMCID： PMC2748124 DOI： 10.1016/j.febslet.2006.09.048
Source DB: PubMed Journal: FEBS Lett ISSN： 0014-5793 Impact factor: 4.124