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Optimized and far-red emitting variants of fluorescent protein eqFP611

Optimized and far-red emitting variants of fluorescent protein eqFP611
Optimized and far-red emitting variants of fluorescent protein eqFP611
Fluorescent proteins (FPs) emitting in the far-red region of the spectrum are highly advantageous for whole-body imaging applications because scattering and absorption of long-wavelength light is markedly reduced in tissue. We characterized variants of the red fluorescent protein eqFP611 with bright fluorescence emission shifted up to 639 nm. The additional red shift is caused by a trans-cis isomerization of the chromophore. The equilibrium between the trans and cis conformations is strongly influenced by amino acid residues 143 and 158. Pseudo monomeric tags were obtained by further genetic engineering. For the red chromophores of eqFP611 variants, molar extinction coefficients of up to 150,000 were determined by an approach that is not affected by the presence of molecules with nonfunctional red chromophores. The bright fluorescence makes the red-shifted eqFP611 variants promising lead structures for the development of near-infrared fluorescent markers. The red fluorescent proteins performed well in cell biological applications, including two-photon imaging.
1074-5521
224-233
Kredel, S.
d2661b60-bdf2-4928-ab60-f21d02cb5f18
Nienhaus, K.
ff9b0d42-5cd8-49a7-8ed4-24db20289091
Wolff, M.
f4474770-dda8-4ed9-8704-1c28569c4e83
Oswald, F.
8fea64d4-21b7-4f41-93dd-0c4ce331c84b
Ivanchenko, S.
99165146-f345-45ef-9f34-e72e8af01c41
Cymer, F.
e9a7d13b-7c8f-42d7-b152-6b8b43766f5c
Jeromin, A.
cbcadb50-e689-48b5-89bc-20a9a5e4f40e
Michel, F.J.
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Spindler, K.D.
6b511593-a50c-40d8-8a8a-bdc6790aaefd
Heilker, R.
8d99f47e-fa07-407f-a649-138d1e6ec789
Nienhaus, G.U.
8f2b40ea-be57-4b32-880f-8a0e5c1585aa
Wiedenmann, J.
ad445af2-680f-4927-90b3-589ac9d538f7
Kredel, S.
d2661b60-bdf2-4928-ab60-f21d02cb5f18
Nienhaus, K.
ff9b0d42-5cd8-49a7-8ed4-24db20289091
Wolff, M.
f4474770-dda8-4ed9-8704-1c28569c4e83
Oswald, F.
8fea64d4-21b7-4f41-93dd-0c4ce331c84b
Ivanchenko, S.
99165146-f345-45ef-9f34-e72e8af01c41
Cymer, F.
e9a7d13b-7c8f-42d7-b152-6b8b43766f5c
Jeromin, A.
cbcadb50-e689-48b5-89bc-20a9a5e4f40e
Michel, F.J.
9d4912bd-e12a-4f1a-b04f-61bf30cfc3ab
Spindler, K.D.
6b511593-a50c-40d8-8a8a-bdc6790aaefd
Heilker, R.
8d99f47e-fa07-407f-a649-138d1e6ec789
Nienhaus, G.U.
8f2b40ea-be57-4b32-880f-8a0e5c1585aa
Wiedenmann, J.
ad445af2-680f-4927-90b3-589ac9d538f7

Kredel, S., Nienhaus, K., Wolff, M., Oswald, F., Ivanchenko, S., Cymer, F., Jeromin, A., Michel, F.J., Spindler, K.D., Heilker, R., Nienhaus, G.U. and Wiedenmann, J. (2008) Optimized and far-red emitting variants of fluorescent protein eqFP611. Chemistry & Biology, 15 (3), 224-233. (doi:10.1016/j.chembiol.2008.02.008).

Record type: Article

Abstract

Fluorescent proteins (FPs) emitting in the far-red region of the spectrum are highly advantageous for whole-body imaging applications because scattering and absorption of long-wavelength light is markedly reduced in tissue. We characterized variants of the red fluorescent protein eqFP611 with bright fluorescence emission shifted up to 639 nm. The additional red shift is caused by a trans-cis isomerization of the chromophore. The equilibrium between the trans and cis conformations is strongly influenced by amino acid residues 143 and 158. Pseudo monomeric tags were obtained by further genetic engineering. For the red chromophores of eqFP611 variants, molar extinction coefficients of up to 150,000 were determined by an approach that is not affected by the presence of molecules with nonfunctional red chromophores. The bright fluorescence makes the red-shifted eqFP611 variants promising lead structures for the development of near-infrared fluorescent markers. The red fluorescent proteins performed well in cell biological applications, including two-photon imaging.

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Published date: 21 March 2008
Organisations: Ocean and Earth Science

Identifiers

Local EPrints ID: 51156
URI: http://eprints.soton.ac.uk/id/eprint/51156
ISSN: 1074-5521
PURE UUID: b445922d-7e53-4203-8a6c-1b42b0d5137d
ORCID for J. Wiedenmann: ORCID iD orcid.org/0000-0003-2128-2943

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Date deposited: 07 May 2008
Last modified: 09 Jan 2022 03:24

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Contributors

Author: S. Kredel
Author: K. Nienhaus
Author: M. Wolff
Author: F. Oswald
Author: S. Ivanchenko
Author: F. Cymer
Author: A. Jeromin
Author: F.J. Michel
Author: K.D. Spindler
Author: R. Heilker
Author: G.U. Nienhaus
Author: J. Wiedenmann ORCID iD

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