Fluorescence of Proteins in 6‐M Guanidine Hydrochloride

作者：P. Pajot · 发表于：European Journal of Biochemistry · 年份：1976 · DOI：10.1111/j.1432-1033.1976.tb10228.x · 被引用次数：209 · 研究领域：Protein Interaction Studies and Fluorescence Analysis、Hemoglobin structure and function、Protein Structure and Dynamics

To determine the tryptophan content in proteins,an analytical ultraviolet fluroescence method is proposed based on making uniform the environment of aromatic chromophores in 6-7 M guanidine hydrochloride. The fluorescence intensity scale is calibrated using standard solutions of free tryptophan. A correlation coefficient between the fluorescence of protein tryptophanyl residues and of free tryptophan was estimated in testing 17 well characterized proteins. This method is particularly suited to proteins carrying groups absorbing in the 290-370 nm region, such as flavin, heme and pyridoxal phosphate and in the presence of substances such as 2-mercaptoethanol which prohibit the use of the spectroscopic or magnetic circular dichroism methods. It is less time-consuming than techniques requiring hydrolysis or chemical reactions.