Infrared Spectroscopy Reveals Multi-step Multi-timescale Photoactivation in the Photoconvertible Protein Archetype Dronpa

Overview

Journal Nat Chem

Specialty Chemistry

Date 2018 Jun 13

PMID 29892029

Citations 15

Authors

Sergey P Laptenok

Agnieszka A Gil

Christopher R Hall

Andras Lukacs

James N Iuliano

Garth A Jones

Gregory M Greetham

Paul Donaldson

Atsushi Miyawaki

Peter J Tonge

Stephen R Meech

Affiliations

Soon will be listed here.

Abstract

Photochromic fluorescent proteins play key roles in super-resolution microscopy and optogenetics. The light-driven structural changes that modulate the fluorescence involve both trans-to-cis isomerization and proton transfer. The mechanism, timescale and relative contribution of chromophore and protein dynamics are currently not well understood. Here, the mechanism of off-to-on-state switching in dronpa is studied using femtosecond-to-millisecond time-resolved infrared spectroscopy and isotope labelling. Chromophore and protein dynamics are shown to occur on multiple timescales, from picoseconds to hundreds of microseconds. Following excitation of the trans chromophore, a ground-state primary product is formed within picoseconds. Surprisingly, the characteristic vibrational spectrum of the neutral cis isomer appears only after several tens of nanoseconds. Further fluctuations in protein structure around the neutral cis chromophore are required to form a new intermediate, which promotes the final proton-transfer reaction. These data illustrate the interplay between chromophore dynamics and the protein environment underlying fluorescent protein photochromism.

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