Probing Variations of Reduction Activity at the Plasma Membrane Using a Targeted Ratiometric FRET Probe

Tarushyam Mukherjee , Sriram Kanvah , Andrey S Klymchenko (1) , Mayeul Collot (1)

1 LBP - Laboratoire de Bioimagerie et Pathologies

Plasma membrane (PM) is the turntable of various reactions that regulate essential functionalities of cells. Among these reactions, the thiol disulfide exchange (TDE) reaction plays an important role in cellular processes. We herein designed a selective probe, called membrane reduction probe (MRP), that is able to report TDE activity at the PM. MRP is based on a green emitting BODIPY PM probe connected to rhodamine through a disulfide bond. MRP is fluorogenic as it is turned off in aqueous media due to aggregation-caused quenching, and once inserted in the PM, it displays a bright red signal due to an efficient fluorescence energy resonance transfer (FRET) between the BODIPY donor and the rhodamine acceptor. In the PM model, the MRP can undergo TDE reaction with external reductive agents as well as with thiolated lipids embedded in the bilayer. Upon TDE reaction, the FRET is turned off and a bright green signal appears allowing a ratiometric readout of this reaction. In cells, the MRP quickly labeled the PM and was able to probe variations of TDE activity using ratiometric imaging. With this tool in hand, we were able to monitor variations of TDE activity at the PM under stress conditions, and we showed that cancer cell lines presented a reduced TDE activity at the PM compared to noncancer cells.

Mots clés: Metrics & More Article Recommendations plasma membrane probe; fluorogenic probe; BODIPY; cell surface reaction; thiol disulfide exchange; FRET; ratiometric imaging

Domaines: Chimie organique

Tarushyam Mukherjee, Sriram Kanvah, Andrey S Klymchenko, Mayeul Collot. Probing Variations of Reduction Activity at the Plasma Membrane Using a Targeted Ratiometric FRET Probe. ACS Applied Materials & Interfaces, 2021, 13, pp.40315 - 40324. ⟨10.1021/acsami.1c11069⟩. ⟨hal-03424429⟩