Department of Chemistry and Research Institute of Basic Sciences, Kyung Hee University, 26 Kyungheedae-ro, Dongdaemun-gu, Seoul 02447, Korea.
Department of Life Science, Chung-Ang University, 84 Heukseok-ro, Dongjak-gu, Seoul 06974, Korea.
J Am Chem Soc. 2020 May 20;142(20):9231-9239. doi: 10.1021/jacs.9b13982. Epub 2020 May 7.
Fluorescence-based amine-reactive dyes are highly valuable for the sensing of amines and the labeling of biomolecules. Although it would be highly desirable, large changes in emission spectra and intensity seldom accompany the conjugation of known amine-reactive dyes to their target molecules. On the contrary, amide bond formation between amines and the pentafluorophenyl () and succinimidyl () esters of -carboxyBODIPY results in significant changes in emission maxima (Δλ: 70-100 nm) and intensity (up to 3000-fold), enabling the fast (down to 5 min) and selective fluorogenic detection and labeling of amines, amino acids, and proteins. This approach further benefits from the demonstrated versatility and high reliability of activated ester chemistry, and background hydrolysis is negligible. The large "turn-on" response is a testament of the extreme sensitivity of carboxyBODIPYs to the minimal changes in electronic properties that distinguish esters from amides. Applications to the detection of food spoilage, staining of proteins on electrophoretic gels or in living cells, and the expedited synthesis of organelle-specific fluorescence microscope imaging agents are further demonstrated.
基于荧光的胺反应染料对于胺的传感和生物分子的标记非常有价值。尽管这是非常理想的，但已知的胺反应染料与它们的目标分子缀合时，发射光谱和强度很少发生大的变化。相反，胺与五氟苯基（）和琥珀酰亚胺基（）酯之间形成酰胺键会导致发射最大值（Δλ：70-100nm）和强度（高达 3000 倍）发生显著变化，从而能够快速（低至 5 分钟）和选择性地检测和标记胺、氨基酸和蛋白质。这种方法进一步受益于已证明的活化酯化学的多功能性和高可靠性，并且背景水解可以忽略不计。大的“开启”响应证明了羧基 BODIPY 对电子性质的微小变化极其敏感，这种变化将酯与酰胺区分开来。进一步证明了其在检测食品变质、电泳凝胶或活细胞中蛋白质染色以及加速细胞器特异性荧光显微镜成像试剂的合成方面的应用。
