动态结合剂交换提高了DNA-PAINT中的蛋白质标记效率,高达15倍
Clemens Steinek1, Isabelle Pachmayr1, Sebastian Strauss1
1Max Planck Institute of Biochemistry, Am Klopferspitz 18, 82152, Martinsried, Germany.
Angewandte Chemie (International ed. in English)
|February 3, 2026
概括
动态绑定器交换 (DyBE) 通过使用可逆绑定器来增强目标采样,改进了DNA-PAINT超分辨率显微镜. 这种方法实现了更高的标记效率,使得蛋白质相互作用的敏感检测,如HER2同位体和EGFR-HER2异位体.
科学领域:
- 生物物理学的生物物理.
- 分子成像学分子成像学
- 细胞生物学 细胞生物学
背景情况:
- DNA-PAINT (纳米级拓图像点积累) 是一种超高分辨率显微镜技术.
- 暂时的,可逆的粘合剂-目标相互作用对于高效的分子成像至关重要.
- 在超分辨率显微镜中,检测具有高偏离率结合剂的目标存在局限性.
研究的目的:
- 引入和验证动态绑定器交换 (DyBE) 作为DNA-PAINT显微镜的增强.
- 提高标签效率,并使以前无法访问的目标能够灵敏地检测.
- 在单分子水平上研究受体蛋白的空间组织,包括HER2和EGFR.
主要方法:
- DyBE利用具有高脱离率的DNA结合结合剂 (例如纳米体) 进行短暂的目标结合.
- 双动力学方案将粘合剂的减速率与DNA-PAINT闪动态相结合.
- 进行了单蛋白分辨率成像,以检测HER2同位体和EGFR-HER2异位体.
主要成果:
- 与传统方法相比,DyBE提高了标签效率高达15倍.
- 能够灵敏地检测出先前存在的HER2同位体和联体诱导的EGFR-HER2异位体.
- 在单个蛋白质水平上展示了受体二分化动态的高保真成像.
结论:
- 通过优化绑定器-目标相互作用,DyBE显著增强了DNA-PAINT超分辨率显微镜.
- 该方法扩大了可用于先进空间蛋白质组学的可用结合剂的目录.
- 通过对受体组织和信号通路进行详细分析,DyBE促进了机械药物研究.
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