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Structural characterization of IrisFP, an optical highlighter undergoing multiple photo-induced transformations

Structural characterization of IrisFP, an optical highlighter undergoing multiple photo-induced transformations
Structural characterization of IrisFP, an optical highlighter undergoing multiple photo-induced transformations
Photoactivatable fluorescent proteins (FPs) are powerful fluorescent highlighters in live cell imaging and offer perspectives for optical nanoscopy and the development of biophotonic devices. Two types of photoactivation are currently being distinguished, reversible photoswitching between fluorescent and nonfluorescent forms and irreversible photoconversion. Here, we have combined crystallography and (in crystallo) spectroscopy to characterize the Phe-173-Ser mutant of the tetrameric variant of EosFP, named IrisFP, which incorporates both types of phototransformations. In its green fluorescent state, IrisFP displays reversible photoswitching, which involves cis–trans isomerization of the chromophore. Like its parent protein EosFP, IrisFP also photoconverts irreversibly to a red-emitting state under violet light because of an extension of the conjugated ?-electron system of the chromophore, accompanied by a cleavage of the polypeptide backbone. The red form of IrisFP exhibits a second reversible photoswitching process, which may also involve cis–trans isomerization of the chromophore. Therefore, IrisFP displays altogether 3 distinct photoactivation processes. The possibility to engineer and precisely control multiple phototransformations in photoactivatable FPs offers exciting perspectives for the extension of the fluorescent protein toolkit.
fluorescent proteins, microspectrophotometry, photoactivation, photochromism, protein crystallography
0027-8424
18343-18348
Adam, Virgile
9576bdff-5f2c-4f0c-a649-e54b43949cab
Lelimousin, Mickaël
02ebb13a-4437-42f1-99ed-8b4fbd963149
Boehme, Susan
61313daa-a907-4072-8b7e-f28c39176cc7
Desfonds, Guillaume
45faafdf-df11-4b9d-aa3c-1894846b6e6a
Nienhaus, Karin
b05bb626-b88d-48d8-b0ed-cc1624de52ca
Field, Martin J.
cce09e58-ee88-4111-ae84-b0d56b1a0da8
Wiedenmann, Joerg
ad445af2-680f-4927-90b3-589ac9d538f7
McSweeney, Sean
97b692ba-083e-4d45-acda-c4aabb8661fa
Nienhaus, G. Ulrich
64eb2ac6-4fa9-416c-a066-f096d79307cb
Bourgeois, Dominique
8ba54f95-5106-47f0-bafb-83fe97c57a9d
Adam, Virgile
9576bdff-5f2c-4f0c-a649-e54b43949cab
Lelimousin, Mickaël
02ebb13a-4437-42f1-99ed-8b4fbd963149
Boehme, Susan
61313daa-a907-4072-8b7e-f28c39176cc7
Desfonds, Guillaume
45faafdf-df11-4b9d-aa3c-1894846b6e6a
Nienhaus, Karin
b05bb626-b88d-48d8-b0ed-cc1624de52ca
Field, Martin J.
cce09e58-ee88-4111-ae84-b0d56b1a0da8
Wiedenmann, Joerg
ad445af2-680f-4927-90b3-589ac9d538f7
McSweeney, Sean
97b692ba-083e-4d45-acda-c4aabb8661fa
Nienhaus, G. Ulrich
64eb2ac6-4fa9-416c-a066-f096d79307cb
Bourgeois, Dominique
8ba54f95-5106-47f0-bafb-83fe97c57a9d

Adam, Virgile, Lelimousin, Mickaël, Boehme, Susan, Desfonds, Guillaume, Nienhaus, Karin, Field, Martin J., Wiedenmann, Joerg, McSweeney, Sean, Nienhaus, G. Ulrich and Bourgeois, Dominique (2008) Structural characterization of IrisFP, an optical highlighter undergoing multiple photo-induced transformations. Proceedings of the National Academy of Sciences, 105 (47), 18343-18348. (doi:10.1073/pnas.0805949105).

Record type: Article

Abstract

Photoactivatable fluorescent proteins (FPs) are powerful fluorescent highlighters in live cell imaging and offer perspectives for optical nanoscopy and the development of biophotonic devices. Two types of photoactivation are currently being distinguished, reversible photoswitching between fluorescent and nonfluorescent forms and irreversible photoconversion. Here, we have combined crystallography and (in crystallo) spectroscopy to characterize the Phe-173-Ser mutant of the tetrameric variant of EosFP, named IrisFP, which incorporates both types of phototransformations. In its green fluorescent state, IrisFP displays reversible photoswitching, which involves cis–trans isomerization of the chromophore. Like its parent protein EosFP, IrisFP also photoconverts irreversibly to a red-emitting state under violet light because of an extension of the conjugated ?-electron system of the chromophore, accompanied by a cleavage of the polypeptide backbone. The red form of IrisFP exhibits a second reversible photoswitching process, which may also involve cis–trans isomerization of the chromophore. Therefore, IrisFP displays altogether 3 distinct photoactivation processes. The possibility to engineer and precisely control multiple phototransformations in photoactivatable FPs offers exciting perspectives for the extension of the fluorescent protein toolkit.

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More information

e-pub ahead of print date: 18 November 2008
Published date: 25 November 2008
Keywords: fluorescent proteins, microspectrophotometry, photoactivation, photochromism, protein crystallography
Organisations: Ocean and Earth Science

Identifiers

Local EPrints ID: 71990
URI: https://eprints.soton.ac.uk/id/eprint/71990
ISSN: 0027-8424
PURE UUID: ad375993-343f-468b-a245-5503b3077971
ORCID for Joerg Wiedenmann: ORCID iD orcid.org/0000-0003-2128-2943

Catalogue record

Date deposited: 13 Jan 2010
Last modified: 12 Nov 2019 01:45

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Contributors

Author: Virgile Adam
Author: Mickaël Lelimousin
Author: Susan Boehme
Author: Guillaume Desfonds
Author: Karin Nienhaus
Author: Martin J. Field
Author: Sean McSweeney
Author: G. Ulrich Nienhaus
Author: Dominique Bourgeois

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