Synthesis and properties of the new thiol-specific reagent difluorescein disulfide: Its application on histone-histone and histone-DNA interactions
✍ Scribed by Edgar Wingender; Alejandro Arellano
- Publisher
- Elsevier Science
- Year
- 1982
- Tongue
- English
- Weight
- 794 KB
- Volume
- 127
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
The synthesis of difluorescein disulfide, a new fluorescent thiol-specific reagent, is described. The absorption and fluorescence spectra and pH dependency of this compound and of several other fluorescein derivatives are presented. Histone H3 labeled with difluorescein disulfide displays a high fluorescence intensity, and conformational changes that affect this parameter could be followed very sensitively. Furthermore, through the cleavage of the labeled histone with dithiothreitol, fluoresceinthiol is obtained which has a very low quantum yield. This fact allows the determination of the kinetics of the cleavage reaction and provides additional information about the environment of the fluorescence probe. The utility of this new labeling reagent and the advantages of both methods for recording fluorescence changes have been demonstrated in the system of histone-histone and histone-DNA interactions.