Archaerhodopsin 3 is an ideal template for the engineering of highly fluorescent optogenetic reporters. - Université de Lille
Article Dans Une Revue Chemical Science Année : 2024

Archaerhodopsin 3 is an ideal template for the engineering of highly fluorescent optogenetic reporters.

Résumé

Archaerhodopsin-3 (AR-3) variants stand out among other rhodopsins in that they display a weak, but voltage-sensitive, near-infrared fluorescence emission. This has led to their application in optogenetics both in cell cultures and small animals. However, in the context of improving the fluorescence characteristics of the next generation of AR-3 reporters, an understanding of their ultrafast light-response in light-adapted conditions, is mandatory. To this end, we present a combined experimental and computational investigation of the excited state dynamics and quantum yields of AR-3 and its DETC and Arch-5 variants. The latter always display a mixture of all-trans/15-anti and 13-cis/15-syn isomers, which leads to a bi-exponential excited state decay. The isomerisation quantum yield is reduced more than 20 times as compared to WT AR-3 and proves that the steady-state fluorescence is induced by a single absorption photon event. In wild-type AR-3, we show that a 300 fs, barrier-less and vibrationally coherent isomerization is driven by an unusual covalent electronic character of its all-trans retinal chromophore leading to a metastable twisted diradical (TIDIR), in clear contrast to the standard charge-transfer scenario established for other microbial rhodopsins. We discuss how the presence of TIDIR makes AR-3 an ideal candidate for the design of variants with a one-photon induced fluorescence possibly reaching the emission quantum yield of the top natural emitter neorhodopsin (NeoR).
Fichier non déposé

Dates et versions

hal-04844689 , version 1 (18-12-2024)

Identifiants

Citer

K. Herasymenko, D. Walisinghe, M. Konno, L. Barneschi, Isabelle de Waele, et al.. Archaerhodopsin 3 is an ideal template for the engineering of highly fluorescent optogenetic reporters.. Chemical Science, 2024, Chem Sci, ⟨10.1039/d4sc05120c⟩. ⟨hal-04844689⟩
0 Consultations
0 Téléchargements

Altmetric

Partager

More