A novel analytical method for in vivo phosphate tracking | Zendy

Gu Hong | Zendy; Lalonde Sylvie | Zendy; Okumoto Sakiko | Zendy; Looger Loren L. | Zendy; Scharff-Poulsen Anne Marie | Zendy; Grossman Arthur R. | Zendy; Kossmann Jens | Zendy; Jakobsen Iver | Zendy; Frommer Wolf B. | Zendy

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A novel analytical method for in vivo phosphate tracking

Author(s) -

Gu Hong,

Lalonde Sylvie,

Okumoto Sakiko,

Looger Loren L.,

Scharff-Poulsen Anne Marie,

Grossman Arthur R.,

Kossmann Jens,

Jakobsen Iver,

Frommer Wolf B.

Publication year - 2006

Publication title -

febs letters

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 1.593

H-Index - 257

eISSN - 1873-3468

pISSN - 0014-5793

DOI - 10.1016/j.febslet.2006.09.048

Subject(s) - förster resonance energy transfer , chemistry , biophysics , fluorescence , phosphate , biochemistry , transporter , biology , gene , physics , quantum mechanics

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 μM 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.

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