A novel analytical method for in vivo phosphate tracking
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A novel analytical method for in vivo phosphate tracking. / Gu, Hong; Lalonde, Sylvie; Okumoto, Sakiko; Looger, Loren L.; Scharff-Poulsen, Anne Marie; Grossman, Arthur R.; Kossmann, Jens; Jakobsen, Iver; Frommer, Wolf B.
I: FEBS Letters, Bind 580, Nr. 25, 2006, s. 5885-5893.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - A novel analytical method for in vivo phosphate tracking
AU - Gu, Hong
AU - Lalonde, Sylvie
AU - Okumoto, Sakiko
AU - Looger, Loren L.
AU - Scharff-Poulsen, Anne Marie
AU - Grossman, Arthur R.
AU - Kossmann, Jens
AU - Jakobsen, Iver
AU - Frommer, Wolf B.
N1 - FLIPPi, fluorescent indicator protein for inorganic phosphate, FRET, fluorescence resonance energy transfer, FP, fluorescent protein
PY - 2006
Y1 - 2006
N2 - 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.
AB - 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.
KW - Former LIFE faculty
KW - Fluorescence energy transfer, Phosphate starvation, Biosensor, Synechococcus
U2 - 10.1016/j.febslet.2006.09.048
DO - 10.1016/j.febslet.2006.09.048
M3 - Journal article
C2 - 17034793
VL - 580
SP - 5885
EP - 5893
JO - F E B S Letters
JF - F E B S Letters
SN - 0014-5793
IS - 25
ER -
ID: 8042551