A photochromic acceptor as a reversible light-driven switch in fluorescence resonance energy transfer (FRET)
A method has been developed for the quantitative determination of fluorescence resonance energy transfer (FRET) based on the modulation of donor fluorescence upon the reversible photoconversion of a photochromic acceptor. A model system was devised, consisting of Lucifer Yellow cadaverine (LYC, dono...
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todo:paper_10106030_v150_n1-3_p177_Song2023-10-03T15:55:59Z A photochromic acceptor as a reversible light-driven switch in fluorescence resonance energy transfer (FRET) Song, L. Jares-Erijman, E.A. Jovin, T.M. Fluorescence microscopy Merocyanine pcFRET Photochromism Spiropyran 1',3' dihydro 1' (2 carboxyethyl) 3,3' dimethyl 6 nitrospiro[2h 1 benzopyran 2,2'(2h) indoline] cadaverine fluorescent dye lucifer yellow merocyanine spiropyran unclassified drug absorption article catalysis concentration (parameters) conjugation controlled study donor energy transfer excitation fluorescence fluorescence microscopy light model molecular interaction molecular stability photochemistry quantitative analysis ultraviolet radiation A method has been developed for the quantitative determination of fluorescence resonance energy transfer (FRET) based on the modulation of donor fluorescence upon the reversible photoconversion of a photochromic acceptor. A model system was devised, consisting of Lucifer Yellow cadaverine (LYC, donor) conjugated to the photochromic molecule, 6-nitroBIPS (1′,3′-dihydro-1′-(2-carboxyethyl)- Near-ultraviolet irradiation catalyzes the conversion of the colorless spiropyran (SP) to the colored merocyanine (MC) form of 6-nitroBIPS. Only the MC form absorbs at the emission wavelengths of the donor, thereby potentiating FRET, as demonstrated by quenching of the donor. Subsequent irradiation in the visible MC absorption band reverts 6-nitroBIPS to the SP form and FRET is inactivated. The acceptor exhibited high photostability under repeated cycles of alternating UV-Vis irradiation. In this model system, the intramolecular FRET efficiency was close to 100%. The observed maximal donor quenching of 34± 3% was indicative of an equilibrium determined by the high quantum efficiency of forward conversion (SP → MC) induced by near-UV irradiation and a low but finite quantum efficiency of the back reaction resulting from excitation of the MC form directly as well as indirectly (by FRET via the donor). A quantitative formalism for the photokinetic scheme was developed. Photochromic FRET (pcFRET) permits repeated, quantitative, and non-destructive FRET determinations for arbitrary relative concentrations of donor and acceptor and thus offers great potential for monitoring dynamic molecular interactions in living cells over extended observation times by fluorescence microscopy. © 2002 Elsevier Science B.V. All rights reserved. Fil:Jares-Erijman, E.A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_10106030_v150_n1-3_p177_Song |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
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Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Fluorescence microscopy Merocyanine pcFRET Photochromism Spiropyran 1',3' dihydro 1' (2 carboxyethyl) 3,3' dimethyl 6 nitrospiro[2h 1 benzopyran 2,2'(2h) indoline] cadaverine fluorescent dye lucifer yellow merocyanine spiropyran unclassified drug absorption article catalysis concentration (parameters) conjugation controlled study donor energy transfer excitation fluorescence fluorescence microscopy light model molecular interaction molecular stability photochemistry quantitative analysis ultraviolet radiation |
| spellingShingle |
Fluorescence microscopy Merocyanine pcFRET Photochromism Spiropyran 1',3' dihydro 1' (2 carboxyethyl) 3,3' dimethyl 6 nitrospiro[2h 1 benzopyran 2,2'(2h) indoline] cadaverine fluorescent dye lucifer yellow merocyanine spiropyran unclassified drug absorption article catalysis concentration (parameters) conjugation controlled study donor energy transfer excitation fluorescence fluorescence microscopy light model molecular interaction molecular stability photochemistry quantitative analysis ultraviolet radiation Song, L. Jares-Erijman, E.A. Jovin, T.M. A photochromic acceptor as a reversible light-driven switch in fluorescence resonance energy transfer (FRET) |
| topic_facet |
Fluorescence microscopy Merocyanine pcFRET Photochromism Spiropyran 1',3' dihydro 1' (2 carboxyethyl) 3,3' dimethyl 6 nitrospiro[2h 1 benzopyran 2,2'(2h) indoline] cadaverine fluorescent dye lucifer yellow merocyanine spiropyran unclassified drug absorption article catalysis concentration (parameters) conjugation controlled study donor energy transfer excitation fluorescence fluorescence microscopy light model molecular interaction molecular stability photochemistry quantitative analysis ultraviolet radiation |
| description |
A method has been developed for the quantitative determination of fluorescence resonance energy transfer (FRET) based on the modulation of donor fluorescence upon the reversible photoconversion of a photochromic acceptor. A model system was devised, consisting of Lucifer Yellow cadaverine (LYC, donor) conjugated to the photochromic molecule, 6-nitroBIPS (1′,3′-dihydro-1′-(2-carboxyethyl)- Near-ultraviolet irradiation catalyzes the conversion of the colorless spiropyran (SP) to the colored merocyanine (MC) form of 6-nitroBIPS. Only the MC form absorbs at the emission wavelengths of the donor, thereby potentiating FRET, as demonstrated by quenching of the donor. Subsequent irradiation in the visible MC absorption band reverts 6-nitroBIPS to the SP form and FRET is inactivated. The acceptor exhibited high photostability under repeated cycles of alternating UV-Vis irradiation. In this model system, the intramolecular FRET efficiency was close to 100%. The observed maximal donor quenching of 34± 3% was indicative of an equilibrium determined by the high quantum efficiency of forward conversion (SP → MC) induced by near-UV irradiation and a low but finite quantum efficiency of the back reaction resulting from excitation of the MC form directly as well as indirectly (by FRET via the donor). A quantitative formalism for the photokinetic scheme was developed. Photochromic FRET (pcFRET) permits repeated, quantitative, and non-destructive FRET determinations for arbitrary relative concentrations of donor and acceptor and thus offers great potential for monitoring dynamic molecular interactions in living cells over extended observation times by fluorescence microscopy. © 2002 Elsevier Science B.V. All rights reserved. |
| format |
JOUR |
| author |
Song, L. Jares-Erijman, E.A. Jovin, T.M. |
| author_facet |
Song, L. Jares-Erijman, E.A. Jovin, T.M. |
| author_sort |
Song, L. |
| title |
A photochromic acceptor as a reversible light-driven switch in fluorescence resonance energy transfer (FRET) |
| title_short |
A photochromic acceptor as a reversible light-driven switch in fluorescence resonance energy transfer (FRET) |
| title_full |
A photochromic acceptor as a reversible light-driven switch in fluorescence resonance energy transfer (FRET) |
| title_fullStr |
A photochromic acceptor as a reversible light-driven switch in fluorescence resonance energy transfer (FRET) |
| title_full_unstemmed |
A photochromic acceptor as a reversible light-driven switch in fluorescence resonance energy transfer (FRET) |
| title_sort |
photochromic acceptor as a reversible light-driven switch in fluorescence resonance energy transfer (fret) |
| url |
http://hdl.handle.net/20.500.12110/paper_10106030_v150_n1-3_p177_Song |
| work_keys_str_mv |
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