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Binding stoichiometry and affinity of fluorescent dyes to proteins in different structural statesстатья
- Авторы: Sulatskaya A.I., Povarova O.I., Kuznetsova I.M., Uversky V.N., Turoverov K.K.
- Сборник: Intrinsically Disordered Proteins: Volume I. Methods and Experimental Tools
- Серия: Methods in Molecular Biology
- Том: 895
- Глава в коллективной монографии
- Год издания: 2012
- Место издания: Humana Press
- Первая страница: 441
- Последняя страница: 460
- Аннотация: Protocol of determination of binding stoichiometry and affinity of fluorescent dyes with proteins in different structural states is proposed. The proposed approach is based on the spectrophotometric determination of concentrations of dye bound to protein and free dye in solutions prepared by equilibrium microdialysis. This technique allows also determining spectral properties of the bound dyes. The restrictions of the use of dye fluorescence intensity for characterization of its interaction with the target protein are discussed. It is shown that the dependence of the dye fluorescence intensity on its optical density together with the data on its binding parameter can give information about the dye fluorescence quantum yield. All procedures are illustrated by interaction of 8-anilino-1-naphthalenesulfonate (ANS) with bovine serum albumin.
- Добавил в систему: Кузнецова Ирина Михайловна