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Photodynamics of red fluorescent proteins studied by fluorescence correlation spectroscopy

Schenk, A., Ivanchenko, S., Röcker, C., Wiedenmann, J. and Nienhaus, G.U. (2004) Photodynamics of red fluorescent proteins studied by fluorescence correlation spectroscopy Biophysical Journal, 86, pp. 384-394.

Record type: Article

Abstract

Red fluorescent proteins are important tools in fluorescence-based life science research. Recently, we have introduced eqFP611, a red fluorescent protein with advantageous properties from the sea anemone Entacmaea quadricolor. Here, we have studied the submillisecond light-driven intramolecular dynamics between bright and dark states of eqFP611 and, for comparison, drFP583 (DsRed) by using fluorescence correlation spectroscopy on protein solutions. A three-state model with one dark and two fluorescent states describes the power-dependence of the flickering dynamics of both proteins at different excitation wavelengths. It involves two light-driven conformational transitions. We have also studied the photodynamics of individual (monomeric) eqFP611 molecules immobilized on surfaces. The flickering rates and dark state fractions of eqFP611 bound to polyethylene glycol-covered glass surfaces were identical to those measured in solution, showing that the bound FPs behaved identically. A second, much slower flickering process was observed on the 10-ms timescale. Deposition of eqFP611 molecules on bare glass surfaces yielded bright fluorescence without any detectable flickering and a >10-fold decreased photobleaching yield. These observations underscore the intimate connection between protein motions and photophysical processes in fluorescent proteins.

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Published date: 2004

Identifiers

Local EPrints ID: 51178
URI: http://eprints.soton.ac.uk/id/eprint/51178
ISSN: 0006-3495
PURE UUID: ac0199fc-ed31-4f03-9d64-1d0cf95b4bef
ORCID for J. Wiedenmann: ORCID iD orcid.org/0000-0003-2128-2943

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Date deposited: 08 May 2008
Last modified: 17 Jul 2017 14:49

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Contributors

Author: A. Schenk
Author: S. Ivanchenko
Author: C. Röcker
Author: J. Wiedenmann ORCID iD
Author: G.U. Nienhaus

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