Kong Xiangxing, Su Fengyu, Zhang Liqiang, Yaron Jordan, Lee Fred, Shi Zhengwei, Tian Yanqing, Meldrum Deirdre R
Center for Biosignatures Discovery Automation, Biodesign Institute, Arizona State University, 1001 S. McAlister Ave., P.O. Box 876501, Tempe, AZ 85287 (USA).
Department of Materials Science and Engineering, South University of Science and Technology of China, No. 1088, Xueyuan Rd., Xili, Nanshan District, Shenzhen, Guangdong, 518055 (China).
Angew Chem Int Ed Engl. 2015 Oct 5;54(41):12053-7. doi: 10.1002/anie.201506038. Epub 2015 Aug 21.
Regulation of intracellular potassium (K(+) ) concentration plays a key role in metabolic processes. So far, only a few intracellular K(+) sensors have been developed. The highly selective fluorescent K(+) sensor KS6 for monitoring K(+) ion dynamics in mitochondria was produced by coupling triphenylphosphonium, borondipyrromethene (BODIPY), and triazacryptand (TAC). KS6 shows a good response to K(+) in the range 30-500 mM, a large dynamic range (Fmax /F0 ≈130), high brightness (ϕf =14.4 % at 150 mM of K(+) ), and insensitivity to both pH in the range 5.5-9.0 and other metal ions under physiological conditions. Colocalization tests of KS6 with MitoTracker Green confirmed its predominant localization in the mitochondria of HeLa and U87MG cells. K(+) efflux/influx in the mitochondria was observed upon stimulation with ionophores, nigericin, or ionomycin. KS6 is thus a highly selective semiquantitative K(+) sensor suitable for the study of mitochondrial potassium flux in live cells.
细胞内钾(K⁺)浓度的调节在代谢过程中起着关键作用。到目前为止,仅开发出了少数几种细胞内K⁺传感器。通过将三苯基鏻、硼二吡咯亚甲基(BODIPY)和三氮杂环壬烷(TAC)偶联,制备了用于监测线粒体中K⁺离子动态的高选择性荧光K⁺传感器KS6。KS6在30 - 500 mM范围内对K⁺表现出良好的响应,具有较大的动态范围(Fmax /F0 ≈130)、高亮度(在150 mM K⁺时ϕf =14.4%),并且在5.5 - 9.0的pH范围内以及生理条件下对其他金属离子不敏感。KS6与MitoTracker Green的共定位测试证实了其在HeLa和U87MG细胞线粒体中的主要定位。在用离子载体尼日利亚菌素或离子霉素刺激后,观察到线粒体中的K⁺外流/内流。因此,KS6是一种高选择性的半定量K⁺传感器,适用于研究活细胞中的线粒体钾通量。