FLUORESCENCE OF DANSYL AMINO ACIDS IN ORGANIC SOLVENTS AND PROTEIN SOLUTIONS
FLUORESCENCE OF DANSYL AMINO ACIDS IN ORGANIC SOLVENTS AND PROTEIN SOLUTIONS
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DOI：
10.1016/0003-9861(67)90526-7
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发表时间：
1967-01-01
影响因子：
3.9
通讯作者：
CHEN, RF
中科院分区：
文献类型：
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作者：
CHEN, RF
丹酰氨基酸在水中仅发出微弱的荧光;校正后的发射光谱的峰在约580 μ m处，量子产率小于0.1。然而，在低介电常数的溶剂中，发射向蓝色移动，量子产率显著增加。因此，在二氧六环中，丹酰DL-色氨酸的荧光在500 μ m处最大，量子产率为0.70。丹磺酰L-脯氨酸在低至3 × 10 ~（-13）摩尔的二氧六环介质中，由于其高的荧光效率和良好的光谱特性，可被荧光光度法测定。丹酰氨基酸结合到具有疏水结合位点的蛋白质，如牛血清白蛋白（BSA）和抹香鲸脱辅基肌红蛋白。同时，荧光向蓝色移动，并且荧光量子产率显著增加。荧光滴定显示，2摩尔丹磺酰L-脯氨酸与每摩尔BSA结合，统计解离常数为2.2 x 10-5 [图片]。血清白蛋白的种属差异反映在白蛋白结合丹酰L-脯氨酸光谱的差异中。与BSA和抹香鲸脱辅基肌红蛋白结合的丹酰氨基酸的荧光的去极化允许计算这些蛋白质的旋转弛豫时间;结合的丹酰氨基酸的荧光寿命在2 X 10-8秒的范围内。丹磺酰氨基酸荧光的定量数据长期缺乏，这些结果对氨基酸分析技术和蛋白质的荧光探针研究具有重要意义。
The dansyl amino acids fluoresce only weakly in water; the peaks of the corrected emission spectra are at about 580 m [mu] and the quantum yields are less than 0.1. However, in solvents of low dielectric constant, the emission shifts toward the blue and the quantum yield increases markedly. Thus in dioxane, the fluorescence of dansyl DL-tryptophan is maximal at 500 m [mu], and the quantum yield is 0.70. Dansyl L-proline can be determined fluoro-metrically in dioxane down to 3 x 10-13 mole as a result of its high fluorescence efficiency and favorable spectral characteristics. Dansyl amino acids bind to proteins having hydrophobic binding sites such as bovine serum albumin (BSA) and sperm whale apomyoglobin. Con-comitantly, the fluorescence shifts toward the blue, and there is a marked increase in fluorescence quantum yield. Fluorometric ti-tration shows that 2 moles of dansyl L-proline bind to each mole of BSA, with a statistical dissociation constant of 2.2 x 10-5 [image]. Species differences in serum albumins are reflected in the differences in the spectra of albumin-bound dansyl L-proline. The depolarization of fluorescence of dansyl amino acids bound to BSA and sperm whale apomyoglobin allows calculation of rotational relaxation times of those proteins; the fluorescence lifetimes of bound dansyl amino acids are in the range of 2 x 10-8 sec. Such quantitative data on dansyl amino acid fluorescence have long been lacking, and these results have implications for amino acid analytical techniques and fluorescent probe studies of proteins.