Identification of the intracellular protein targets of a bio-active clickable half-sandwich iridium complex by chemical proteomics

Robin Ramos (1, 2, 3) , Anthi Karaiskou (4, 3) , Candice Botuha (1, 2, 3) , Michaël Trichet (4, 3) , Florent Dingli (5) , Jérémy Forté (6, 3, 7, 1, 8, 9) , France Lam (3) , Alexis Canette (3) , Chloé Chaumeton (3) , Murielle Salome (10) , Thomas Chenuel (3, 11) , Céline Bergonzi (11, 3) , Philippe Meyer (11, 3) , Sylvain Bohic (12) , Damarys Loew (5) , Michèle Salmain (1, 2, 3) , Joëlle Sobczak-Thépot (4, 3)

1 IPCM - Institut Parisien de Chimie Moléculaire
2 CHEMBIO - Chemical Biology
3 SU - Sorbonne Université
4 CRSA - Centre de Recherche Saint-Antoine
5 Institut Curie [Paris]
6 INC-CNRS - Institut de Chimie - CNRS Chimie
7 UFR de Chimie - UFR 926 [Sorbonne Université]
8 FR 2769 - Chimie Moléculaire de Paris Centre
9 CNRS - Centre National de la Recherche Scientifique
10 ESRF - European Synchrotron Radiation Facility [Grenoble]
11 LBMCE - Laboratoire de Biologie Moléculaire et Cellulaire des Eucaryotes
12 STROBE - Synchrotron Radiation for Biomedicine = Rayonnement SynchroTROn pour la Recherche BiomédicalE

Abstract Identification of intracellular targets of anticancer drug candidates provides key information on their mechanism of action. Exploiting the ability of the anticancer (C^N)-chelated half-sandwich iridium(III) complexes to covalently bind proteins, click chemistry with a bioorthogonal azido probe was used to localize a phenyloxazoline-chelated iridium complex within cells and profile its interactome at the proteome-wide scale. Proteins involved in protein folding and actin cytoskeleton regulation were identified as high affinity targets. Upon iridium complex treatment, HSP90 folding activity was inhibited in vitro and major cytoskeleton disorganization was observed. We used a wide array of imaging and biochemical methods to validate selected targets and obtain a multiscale overview of the effects of this complex on live human cells. We demonstrate that it behaves as a dual agent, inducing both electrophilic and oxidative stresses in cells that account for its cytotoxicity.

Domains: Chemical Sciences; Life Sciences [q-bio]

Robin Ramos, Anthi Karaiskou, Candice Botuha, Michaël Trichet, Florent Dingli, et al.. Identification of the intracellular protein targets of a bio-active clickable half-sandwich iridium complex by chemical proteomics. 2023. ⟨hal-04284979⟩