Photocontrol of Protein Activity in Cultured Cells and Zebrafish with One‐ and Two‐Photon Illumination

• DOI: 10.1002/cbic.201000008
• Publication Date: 2010-02-24T17:26:29Z
• Authors: Deepak Kumar Sinha; Pierre Neveu; Nathalie Gagey; Isabelle Aujard; Chouaha Benbrahim...; Thomas Le Saux; Christine Rampon; Carole Gauron; Bernard Goetz; Sylvie Dubruille; Marc Baaden; Michel Volovitch; David Bensimon; Sophie Vriz; Ludovic Jullien
• Keywords: Inducer

We have implemented a noninvasive optical method for the fast control of protein activity in live zebrafish embryo. It relies on releasing fused to modified estrogen receptor ligand binding domain from its complex with cytoplasmic chaperones, upon local photoactivation nonendogenous caged inducer. Molecular dynamics simulations were used design cyclofen-OH, photochemically stable inducer specific 4-hydroxy-tamoxifen (ER(T2)). Cyclofen-OH was easily synthesized two steps good yields. At submicromolar concentrations, it activates proteins ER(T2) receptor. This shown cultured cells and embryos through emission properties subcellular localization properly engineered fluorescent proteins. successfully various photolabile protecting groups. One particular compound efficient photoinducing nuclear translocation either globally (with 365 nm UV illumination) or locally focused laser two-photon illumination at 750 nm). The present photocontrol could be more generally investigate important physiological processes (e.g., embryogenesis, organ regeneration carcinogenesis) high spatiotemporal resolution.