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 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.
Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
