基因编码素逆转染色体电荷并使酸性囊泡中的光蛋白亮化
Caiyun Fu1, Tomonori Kobayashi, Nanxi Wang
1College of Life Sciences , Zhejiang Sci-Tech University , Hangzhou , China.
Journal of the American Chemical Society
|August 23, 2018
概括
研究人员开发了一种新的酸亮光蛋白 (abFP),在酸性环境中发光,与标准光蛋白不同. 这种创新允许清晰的图像酸性有机体如 lysosomes,克服以前的背景光问题.
科学领域:
- 细胞生物学
- 生物化学
- 分子成像
背景情况:
- 酸性囊泡和细胞器对于细胞内细胞分裂和降解等过程至关重要.
- 常规的光蛋白在酸性pH下通常是非光的,阻碍了这些细胞器的可视化.
- 现有的光报告器会引起背景噪声,使酸性细胞区的研究复杂化.
研究的目的:
- 开发一种在酸性条件下特别明亮的新型光蛋白.
- 在成像酸性细胞环境中克服现有的光蛋白的局限性.
- 能够对酸性囊泡和器官中的分子进行时空分辨率研究.
主要方法:
- 通过将含素的氨基酸 (Qui) 纳入EGFP染色体,设计了一种酸亮化光蛋白 (abFP).
- 修改了染色体的电荷特性,以在酸性pH下实现光增加.
- 在实验室,大肠杆菌和哺乳动物细胞中验证了abFP的性能,包括在溶酶体中对abFP标记的受体的成像.
主要成果:
- 新型abFP在酸性pH下表现出强烈的光,而在中性pH下则非光,从而扭转了典型的pH-光特征.
- 的结合导致了阴离子染色体,导致了pH依赖的光.
- 与EGFP标记的蛋白质不同,abFP标记的蛋白质在溶酶体中成功成像,没有背景干扰.
结论:
- 基染色体策略为开发特定酸的光探针提供了一种新方法.
- abFPs提供了一个强大的工具来研究与最小背景的酸性囊泡和有机体相关的分子.
- 这种策略可以扩展到各种模型系统中创建高级酸性成像的颜色调色板.
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