Department of Chemistry and the Howard Hughes Medical Institute, University of California, Berkeley, California 94720, USA.
J Am Chem Soc. 2010 Feb 3;132(4):1194-5. doi: 10.1021/ja907778b.
We present the synthesis, properties, and biological applications of Ratio-Coppersensor-1 (RCS1), a new water-soluble fluorescent sensor for ratiometric imaging of copper in living cells. RCS1 combines an asymmetric BODIPY reporter and thioether-based ligand receptor to provide high selectivity and sensitivity for Cu(+) over other biologically relevant metal ions, including Cu(2+) and Zn(2+), a ca. 20-fold fluorescence ratio change upon Cu(+) binding, and visible excitation and emission profiles compatible with standard fluorescence microscopy filter sets. Live-cell confocal microscopy experiments show that RCS1 is membrane-permeable and can sense changes in the levels of labile Cu(+) pools within living cells by ratiometric imaging, including expansion of endogenous stores of exchangeable intracellular Cu(+) triggered by ascorbate stimulation in kidney and brain cells.
我们介绍了 Ratio-Coppersensor-1（RCS1）的合成、性质和生物学应用，这是一种新的水溶性荧光传感器，可用于活细胞中铜的比色成像。RCS1 结合了不对称的 BODIPY 报告基团和硫醚基配体受体，为 Cu(+)提供了高选择性和灵敏度，超过了其他生物相关的金属离子，包括 Cu(2+)和 Zn(2+)，Cu(+)结合后荧光比率变化约为 20 倍，并且可见激发和发射光谱与标准荧光显微镜滤光片兼容。活细胞共聚焦显微镜实验表明，RCS1 具有膜透过性，可以通过比色成像来检测活细胞中不稳定 Cu(+)池的水平变化，包括抗坏血酸刺激在肾和脑细胞中引发的内源性可交换细胞内 Cu(+)储存的扩张。
