下一代色到远红色的可光转换光蛋白用于单分子显微镜和蛋白质动态跟踪
Oksana M Subach1, Fakorede Olumayowa2,3,4, Muyuan Tang3
1Complex of NBICS Technologies, National Research Center "Kurchatov Institute", 123182, Moscow, Russia.
Scientific reports
|November 17, 2025
概括
介绍PSmOrange3,一种新型的可光转换光蛋白,可以在绿光下最大限度地减少不必要的转换,从而实现精确的蛋白质跟踪和超高分辨率成像. 这种先进的变体为蜂应用提供了增强的亮度和光稳定性.
科学领域:
- 生物化学 生物化学
- 生物物理学的生物物理.
- 细胞生物学 细胞生物学
背景情况:
- 像PSmOrange和PSmOrange2这样的可光转换光蛋白 (FP) 对先进的成像技术至关重要.
- 然而,它们在激发光下进行光转换的倾向使细胞成像复杂化.
- 需要FP具有更好的特异性和性能.
研究的目的:
- 开发下一代PSmOrange变种,PSmOrange3,在绿光下进行最小的光转换和高光转换对比度.
- 描述PSmOrange3.3的光物理特性,亮度和光稳定性.
- 验证PSmOrange3在活细胞和固定细胞成像中的实用性,包括光激活局部化显微镜 (PALM).
主要方法:
- 结晶学以确定PSmOrange的结构3.
- 质谱法用于分析光转换前后的染色体结构.
- 在细胞环境中进行光转换和光测量.
- 光激活局部化显微镜 (PALM) 用于超高分辨率的图像处理.
主要成果:
- PSmOrange3在紫蓝光 (430-470 nm) 中表现出高效的色到远红色的光转换,在绿光 (550-570 nm) 中表现出最小的光转换.
- 在色的形式中,PSmOrange3比PSmOrange2.2更明亮,更稳定.
- 晶体结构揭示了导致PSmOrange3改善性能的突变.
- PSmOrange3在活体和固定哺乳动物细胞中成功地表现出局部化和光转换,在PALM成像中实现了高精度.
结论:
- PSmOrange3是一种优质的可光转换光蛋白,具有增强的特异性和亮度.
- 它克服了之前PSmOrange变体的局限性,减少了细胞成像中的工件.
- PSmOrange3是蛋白质跟踪,超分辨率显微镜和其他先进的活细胞成像应用程序的宝贵工具.
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