Unknown

Dataset Information

0

Anomalous negative fluorescence anisotropy in yellow fluorescent protein (YFP 10C): quantitative analysis of FRET in YFP dimers.


ABSTRACT: Yellow fluorescent protein (YFP) is widely used as a genetically encoded fluorescent marker in biology. In the course of a comprehensive study of this protein, we observed an unusual, negative fluorescence anisotropy at pH 6.0 (McAnaney, T. B., Zeng, W., Doe, C. F. E., Bhanji, N., Wakelin, S., Pearson, D. S., Abbyad, P., Shi, X., Boxer, S. G., and Bagshaw, C. R. (2005) Biochemistry 44, 5510-5524). Here we report that the fluorescence anisotropy of YFP 10C depends on protein concentration in the low micromolar range that was not expected. We propose that the negative anisotropy is a result of unidirectional Förster resonance energy transfer (FRET) in a dimer of YFP, with the donor chromophore in the neutral form and the acceptor chromophore in the anionic form. This unusual mechanism is supported by studies of a monomeric YFP (A206K YFP) and transient-absorption spectroscopy of YFP 10C. A detailed analysis of the chromophore transition dipole moment direction is presented. The anisotropy and rate constant of this energy transfer are consistent with values produced by an analysis of the dimer structure observed in crystals.

SUBMITTER: Shi X 

PROVIDER: S-EPMC2570256 | biostudies-literature | 2007 Dec

REPOSITORIES: biostudies-literature

altmetric image

Publications

Anomalous negative fluorescence anisotropy in yellow fluorescent protein (YFP 10C): quantitative analysis of FRET in YFP dimers.

Shi Xinghua X   Basran Jaswir J   Seward Harriet E HE   Childs William W   Bagshaw Clive R CR   Boxer Steven G SG  

Biochemistry 20071121 50


Yellow fluorescent protein (YFP) is widely used as a genetically encoded fluorescent marker in biology. In the course of a comprehensive study of this protein, we observed an unusual, negative fluorescence anisotropy at pH 6.0 (McAnaney, T. B., Zeng, W., Doe, C. F. E., Bhanji, N., Wakelin, S., Pearson, D. S., Abbyad, P., Shi, X., Boxer, S. G., and Bagshaw, C. R. (2005) Biochemistry 44, 5510-5524). Here we report that the fluorescence anisotropy of YFP 10C depends on protein concentration in the  ...[more]

Similar Datasets

| S-EPMC5309863 | biostudies-literature
| S-EPMC2718156 | biostudies-literature
| S-EPMC5533704 | biostudies-literature
| S-EPMC7840444 | biostudies-literature
| S-EPMC4523203 | biostudies-literature
| S-EPMC4215982 | biostudies-literature
| S-EPMC3340606 | biostudies-literature
| S-EPMC2586569 | biostudies-literature
| S-EPMC2877369 | biostudies-literature
| S-EPMC7699296 | biostudies-literature