Dark proteins disturb multichromophore coupling in tetrameric fluorescent proteins

Christian Blum, Alfred J. Meixner, Vinod Subramaniam

Research output: Contribution to journalArticleAcademicpeer-review

5 Citations (Scopus)

Abstract

DsRed is representative of the tetrameric reef coral fluorescent proteins that constitute particularly interesting coupled multichromophoric systems. Either a green emitting or a red emitting chromophore can form within each of the monomers of the protein tetramer. Within the tetramers the chromophores are thought to be efficiently fluorescence resonance energy transfer (FRET) coupled. We have used spectrally resolved room temperature single molecule spectroscopy to address the issue of FRET and the role of dark proteins within single protein tetramers of DsRed and its variants DsRed2, DsRed_N42H and AG4. Our results show that for the majority of the tetramers the different chromophores are indeed effectively coupled. However, in a fraction of the tetramers that is characteristic for each DsRed variant analyzed, we observe a lack of effective FRET coupling. For these tetramers we invoke the existence of dark proteins lacking a functional chromophore that interrupt the energy transfer chain within the multichromophoric system. We show that these species lead to donor dequenching that strongly influences the bulk emission spectra.
Original languageUndefined
Pages (from-to)114-121
JournalJournal of biophotonics
Volume4
Issue number1-2
DOIs
Publication statusPublished - 2011

Keywords

  • METIS-272228
  • IR-104423

Cite this

Blum, Christian ; Meixner, Alfred J. ; Subramaniam, Vinod. / Dark proteins disturb multichromophore coupling in tetrameric fluorescent proteins. In: Journal of biophotonics. 2011 ; Vol. 4, No. 1-2. pp. 114-121.
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abstract = "DsRed is representative of the tetrameric reef coral fluorescent proteins that constitute particularly interesting coupled multichromophoric systems. Either a green emitting or a red emitting chromophore can form within each of the monomers of the protein tetramer. Within the tetramers the chromophores are thought to be efficiently fluorescence resonance energy transfer (FRET) coupled. We have used spectrally resolved room temperature single molecule spectroscopy to address the issue of FRET and the role of dark proteins within single protein tetramers of DsRed and its variants DsRed2, DsRed_N42H and AG4. Our results show that for the majority of the tetramers the different chromophores are indeed effectively coupled. However, in a fraction of the tetramers that is characteristic for each DsRed variant analyzed, we observe a lack of effective FRET coupling. For these tetramers we invoke the existence of dark proteins lacking a functional chromophore that interrupt the energy transfer chain within the multichromophoric system. We show that these species lead to donor dequenching that strongly influences the bulk emission spectra.",
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author = "Christian Blum and Meixner, {Alfred J.} and Vinod Subramaniam",
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Dark proteins disturb multichromophore coupling in tetrameric fluorescent proteins. / Blum, Christian; Meixner, Alfred J.; Subramaniam, Vinod.

In: Journal of biophotonics, Vol. 4, No. 1-2, 2011, p. 114-121.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Dark proteins disturb multichromophore coupling in tetrameric fluorescent proteins

AU - Blum, Christian

AU - Meixner, Alfred J.

AU - Subramaniam, Vinod

PY - 2011

Y1 - 2011

N2 - DsRed is representative of the tetrameric reef coral fluorescent proteins that constitute particularly interesting coupled multichromophoric systems. Either a green emitting or a red emitting chromophore can form within each of the monomers of the protein tetramer. Within the tetramers the chromophores are thought to be efficiently fluorescence resonance energy transfer (FRET) coupled. We have used spectrally resolved room temperature single molecule spectroscopy to address the issue of FRET and the role of dark proteins within single protein tetramers of DsRed and its variants DsRed2, DsRed_N42H and AG4. Our results show that for the majority of the tetramers the different chromophores are indeed effectively coupled. However, in a fraction of the tetramers that is characteristic for each DsRed variant analyzed, we observe a lack of effective FRET coupling. For these tetramers we invoke the existence of dark proteins lacking a functional chromophore that interrupt the energy transfer chain within the multichromophoric system. We show that these species lead to donor dequenching that strongly influences the bulk emission spectra.

AB - DsRed is representative of the tetrameric reef coral fluorescent proteins that constitute particularly interesting coupled multichromophoric systems. Either a green emitting or a red emitting chromophore can form within each of the monomers of the protein tetramer. Within the tetramers the chromophores are thought to be efficiently fluorescence resonance energy transfer (FRET) coupled. We have used spectrally resolved room temperature single molecule spectroscopy to address the issue of FRET and the role of dark proteins within single protein tetramers of DsRed and its variants DsRed2, DsRed_N42H and AG4. Our results show that for the majority of the tetramers the different chromophores are indeed effectively coupled. However, in a fraction of the tetramers that is characteristic for each DsRed variant analyzed, we observe a lack of effective FRET coupling. For these tetramers we invoke the existence of dark proteins lacking a functional chromophore that interrupt the energy transfer chain within the multichromophoric system. We show that these species lead to donor dequenching that strongly influences the bulk emission spectra.

KW - METIS-272228

KW - IR-104423

U2 - 10.1002/jbio.201000075

DO - 10.1002/jbio.201000075

M3 - Article

VL - 4

SP - 114

EP - 121

JO - Journal of biophotonics

JF - Journal of biophotonics

SN - 1864-063X

IS - 1-2

ER -