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Abstract
Tryptophan is a fluorescent amino acid that is commonly used as a probe in protein structure-dynamics studies. It is also widely used as a convenient standard for measurements of fluorescence quantum yields of organic and bioorganic molecules. Literature values of the fluorescence quantum yield range from 0.13 to 0.15 upon excitation of tryptophan at 270 nm in aqueous solution. Even though the fluorescence quantum yield of tryptophan is known with a high degree of accuracy upon excitation at 270 nm (i.e., ca. ± 0.02), this value is being used indiscriminately in the literature for the determination of the fluorescence quantum yield of molecules at other excitation wavelengths. In those studies, the assumption has been made that the magnitude of the fluorescence quantum yield of tryptophan does not change with excitation wavelength. This assumption is based on the so-called Kasha’s rule, which states that fluorescence emission occurs in appreciable yield only from the lowest excited singlet state of an organic molecule. Consequently, the fluorescence quantum yield is typically independent of the excitation wavelength used. However, many molecules are known to not obey the Kasha’s rule. Therefore, in this study, we examine the hypothesis that the fluorescence quantum yield of tryptophan does not depend on the excitation wavelength used. We also investigate if the fluorescence quantum yield of tryptophan varies when the salt concentration of the phosphate buffer solution at a physiological pH of 7.4 is varied. The results demonstrate that the fluorescence quantum yield of tryptophan depends on excitation wavelength. It is, however, independent of the ionic strength of the buffered solution in a salt concentration range from 5 to 100 millimolar. A photophysical relaxation mechanism that satisfactorily explains the excitation wavelength dependence of the fluorescence quantum yield of tryptophan will be presented.
Symposium Date
Fall 12-1-2012
Keywords
tryptophan, fluorescence, quantum yield
Disciplines
Chemistry
Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License
Recommended Citation
Abbass, Nadia; Ortiz-Rodríguez, Luis A.; and Crespo-Hernández, Carlos E., "Is the Fluorescence Quantum Yield of Tryptophan Independent of Excitation Wavelength?" (2012). Intersections Fall 2020. 3.
https://commons.case.edu/intersections-fa20/3