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

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TitrePhotocontrol of Protein Activity in Cultured Cells and Zebrafish with One- and Two-Photon Illumination
Type de publicationJournal Article
Nouvelles publications2010
AuteursSinha, DK, Neveu P, Gagey N, Aujard I, Benbrahim-Bouzidi C, Le Saux T, Rampon C, Gauron C, Goetz B, Dubruille S, Baaden M, Volovitch M, Bensimon D, Vriz S, Jullien L
JournalChembiochem
Volume11
Fascicule5
Pagination653-663
Année de publicationMar
Numéro1439-4227
Accession NumberISI:000276536000008
Résumé

We have implemented a noninvasive optical method for the fast control of protein activity in a live zebrafish embryo. It relies on releasing a protein fused to a modified estrogen receptor ligand binding domain from its complex with cytoplasmic chaperones, upon the local photoactivation of a nonendogenous caged inducer. Molecular dynamics simulations were used to design cyclofen-OH, a photochemically stable inducer, of the receptor specific for 4-hydroxy-tamoxifen (ERT2). Cyclofen-OH was easily synthesized in two steps with good yields. At submicromolar concentrations, it activates proteins fused to the ERT2 receptor. This was shown in cultured cells and in zebrafish embryos through emission properties and subcellular localization of properly engineered fluorescent proteins. Cyclofen-OH was successfully caged with various photolabile protecting groups. One particular caged compound was efficient in photoinducing the nuclear translocation of fluorescent proteins either globally (with 365 nm UV illumination) or locally (with a focused UV laser or with two-photon illumination at 750 nm). The present method for photocontrol of protein activity could be used more generally to investigate important physiological processes (e.g., in embryogenesis, organ regeneration and carcinogenesis) with high spatiotemporal resolution.

URL<Go to ISI>://000276536000008
DOI10.1002/cbic.201000008
Importer un fichierPhotocontrol of Protein Activity in Cultured Cells and Zebrafish with One- and Two-Photon Illumination
Unité de rattachement: 
UMR 8640