Lee Jae-Jung, Son Jyunghyun, Ha Hyung-Ho, Chang Young-Tae
Laboratory of Bioimaging Probe Development, Singapore Bioimaging Consortium, Agency for Science, Technology and Research, Biopolis, Singapore.
Mol Biosyst. 2011 Apr;7(4):1270-6. doi: 10.1039/c0mb00327a. Epub 2011 Feb 10.
Imaging a specific protein of interest in live cell has versatile applications in biological research. Recently, diverse chemical tags have been developed to overcome the limits of autofluorescence protein (FP) tags. However, current chemical methods still need to be progressed to compete with FPs in the scope of specificity and convenience in staining procedure. We report a novel protein tagging method that provides a convenient and specific fluorescent labeling of membrane proteins. Ω tag is developed by employing a mammalian enzyme glutathione sulfur-transferase omega 1 (GSTO1) and its partner dye, 5-bromomethyl fluorescein (BMF). Ω-tagged membrane proteins were labeled by BMF efficiently for live cell imaging and in-gel analysis. Endocytosis of epidermal growth factor receptor (EGFR) was successfully visualized by using this Ω tagging system. Ω tag will provide a convenient tool for the physiological study of membrane proteins in live cells.
对活细胞中特定的目标蛋白质进行成像在生物学研究中具有广泛的应用。最近，人们开发了多种化学标签来克服自发荧光蛋白（FP）标签的局限性。然而，目前的化学方法在染色过程的特异性和便利性方面仍需改进，以与荧光蛋白竞争。我们报道了一种新型的蛋白质标记方法，该方法可为膜蛋白提供便捷且特异的荧光标记。Ω标签是通过利用哺乳动物酶谷胱甘肽硫转移酶ω1（GSTO1）及其伴侣染料5-溴甲基荧光素（BMF）开发而成。带有Ω标签的膜蛋白可被BMF高效标记，用于活细胞成像和凝胶内分析。利用这种Ω标记系统成功地可视化了表皮生长因子受体（EGFR）的内吞作用。Ω标签将为活细胞中膜蛋白的生理学研究提供一种便捷的工具。
