A reassessment of synchronous fluorescence in the separation of Trp and Tyr contributions in protein emission and in the determination of conformational changes

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

A reassessment of synchronous fluorescence in the separation of Trp and Tyr contributions in protein emission and in the determination of conformational changes. / Bobone, Sara; van de Weert, Marco; Stella, Lorenzo.

In: Journal of Molecular Structure: THEOCHEM, Vol. 1077, 05.12.2014, p. 68-76.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Bobone, S, van de Weert, M & Stella, L 2014, 'A reassessment of synchronous fluorescence in the separation of Trp and Tyr contributions in protein emission and in the determination of conformational changes', Journal of Molecular Structure: THEOCHEM, vol. 1077, pp. 68-76. https://doi.org/10.1016/j.molstruc.2014.01.004

APA

Bobone, S., van de Weert, M., & Stella, L. (2014). A reassessment of synchronous fluorescence in the separation of Trp and Tyr contributions in protein emission and in the determination of conformational changes. Journal of Molecular Structure: THEOCHEM, 1077, 68-76. https://doi.org/10.1016/j.molstruc.2014.01.004

Vancouver

Bobone S, van de Weert M, Stella L. A reassessment of synchronous fluorescence in the separation of Trp and Tyr contributions in protein emission and in the determination of conformational changes. Journal of Molecular Structure: THEOCHEM. 2014 Dec 5;1077:68-76. https://doi.org/10.1016/j.molstruc.2014.01.004

Author

Bobone, Sara ; van de Weert, Marco ; Stella, Lorenzo. / A reassessment of synchronous fluorescence in the separation of Trp and Tyr contributions in protein emission and in the determination of conformational changes. In: Journal of Molecular Structure: THEOCHEM. 2014 ; Vol. 1077. pp. 68-76.

Bibtex

@article{2bd7fdb4cec84557a898d3714548014b,
title = "A reassessment of synchronous fluorescence in the separation of Trp and Tyr contributions in protein emission and in the determination of conformational changes",
abstract = "Synchronous fluorescence spectra are performed by simultaneously scanning both the excitation and emission wavelengths, and are widely used to analyze complex mixtures of fluorophores, since they yield narrower bands than traditional excitation or emission spectra. Many recent studies claim that synchronous spectra are able to separate tryptophan (Trp) and tyrosine (Tyr) emission in proteins, and use this approach to analyze conformational transitions induced by ligand binding. Here, the reliability of this method is reassessed, studying mixtures of the two intrinsic protein fluorophores in different solvents, as well as a real protein (bovine serum albumin). Unfortunately, synchronous spectra were found to be unreliable in the separation of Trp and Tyr emission components in proteins. A simple alternative approach based on the deconvolution of emission spectra is presented. In addition, an equation predicting the synchronous spectrum of a specific fluorophore from its excitation and emission spectra has been derived.",
keywords = "Conformational transitions, Fluorescence spectroscopy, Ligand binding, Proteins, Spectral deconvolution, Synchronous fluorescence",
author = "Sara Bobone and {van de Weert}, Marco and Lorenzo Stella",
year = "2014",
month = dec,
day = "5",
doi = "10.1016/j.molstruc.2014.01.004",
language = "English",
volume = "1077",
pages = "68--76",
journal = "Computational and Theoretical Chemistry",
issn = "2210-271X",
publisher = "Elsevier",

}

RIS

TY - JOUR

T1 - A reassessment of synchronous fluorescence in the separation of Trp and Tyr contributions in protein emission and in the determination of conformational changes

AU - Bobone, Sara

AU - van de Weert, Marco

AU - Stella, Lorenzo

PY - 2014/12/5

Y1 - 2014/12/5

N2 - Synchronous fluorescence spectra are performed by simultaneously scanning both the excitation and emission wavelengths, and are widely used to analyze complex mixtures of fluorophores, since they yield narrower bands than traditional excitation or emission spectra. Many recent studies claim that synchronous spectra are able to separate tryptophan (Trp) and tyrosine (Tyr) emission in proteins, and use this approach to analyze conformational transitions induced by ligand binding. Here, the reliability of this method is reassessed, studying mixtures of the two intrinsic protein fluorophores in different solvents, as well as a real protein (bovine serum albumin). Unfortunately, synchronous spectra were found to be unreliable in the separation of Trp and Tyr emission components in proteins. A simple alternative approach based on the deconvolution of emission spectra is presented. In addition, an equation predicting the synchronous spectrum of a specific fluorophore from its excitation and emission spectra has been derived.

AB - Synchronous fluorescence spectra are performed by simultaneously scanning both the excitation and emission wavelengths, and are widely used to analyze complex mixtures of fluorophores, since they yield narrower bands than traditional excitation or emission spectra. Many recent studies claim that synchronous spectra are able to separate tryptophan (Trp) and tyrosine (Tyr) emission in proteins, and use this approach to analyze conformational transitions induced by ligand binding. Here, the reliability of this method is reassessed, studying mixtures of the two intrinsic protein fluorophores in different solvents, as well as a real protein (bovine serum albumin). Unfortunately, synchronous spectra were found to be unreliable in the separation of Trp and Tyr emission components in proteins. A simple alternative approach based on the deconvolution of emission spectra is presented. In addition, an equation predicting the synchronous spectrum of a specific fluorophore from its excitation and emission spectra has been derived.

KW - Conformational transitions

KW - Fluorescence spectroscopy

KW - Ligand binding

KW - Proteins

KW - Spectral deconvolution

KW - Synchronous fluorescence

U2 - 10.1016/j.molstruc.2014.01.004

DO - 10.1016/j.molstruc.2014.01.004

M3 - Journal article

AN - SCOPUS:84907672923

VL - 1077

SP - 68

EP - 76

JO - Computational and Theoretical Chemistry

JF - Computational and Theoretical Chemistry

SN - 2210-271X

ER -

ID: 125559773