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开发HaloTag和SNAP-Tag化学诱导体的二聚化以探测受体寡聚化和下游信号传输

Michael Trumpp^1,2, Blaise Gatin-Fraudet³, Kjell Bruckmann¹

¹Institute of Chemistry and Biochemistry - Biochemistry, Freie Universität Berlin, Berlin, Germany.

Angewandte Chemie (International ed. in English)

|June 30, 2025

概括

这项研究为HaloTag和SNAP-Tag蛋白质引入了新的化学二元化诱导剂 (CID),从而能够精确控制蛋白质相互作用. 这些工具帮助研究人员通过诱导和验证受体二分化来理解复杂的信号通路.

科学领域:

分子生物学分子生物学
细胞信号传递细胞信号传递
生物化学生物化学

背景情况:

蛋白质与蛋白质之间的相互作用对于细胞信号通路至关重要,特别是那些由连接体与受体结合启动的信号通路.
使用抗体用于蛋白质复合体诱导的传统方法在石化计,几何控制和特异性方面存在局限性.
化学二元化诱导剂 (CID) 提供了一种精确且共价的方法来控制蛋白质的接近性.

研究的目的:

扩大用于HaloTag (Halo) 和SNAP-Tag (SNAP) 融合蛋白的化学二元化诱导剂 (CID) 的工具包.
开发和验证基于Förster共振能量转移 (FRET) 的传感器,用于诱导和验证蛋白质接近.
通过使用新型CID来研究细胞外受体的连接体独立二分化.

主要方法:

开发新的基 (BG) 和HaloTag连接体 (HTL) 交叉链接剂,具有不同长度的聚乙烯糖醇链接剂.
集成基于FRET的传感器来监测和确认诱导的蛋白质二分化.
将CID应用于细胞外标记的TGFβ,BMP,神经营养因子和转基因型谷氨酸受体.

主要成果:

成功扩展CID工具包用于Halo和SNAP融合蛋白.
使用基于FRET的传感器证明诱导和验证的蛋白质接近性.
在各种细胞外受体上建立CID,使得可以研究连接体独立的异构分子二元化.

结论:

与传统的基于抗体的方法相比,开发的CID提供了更高的精度和对蛋白质-蛋白质相互作用的控制.
基于FRET的传感器提供了一种可靠的方法,可以实时诱导和验证蛋白质的接近性.
这种扩展的CID工具包有助于阐明由连接体独立受体二分化驱动的信号机制.

关键词:

在 BMP/TGFβ 和 TrkB 信号中.分解的化学诱导剂.在FRET传感器上.在HaloTag中使用HaloTag.蛋白质的近距离关系受体的寡合化受体.这是SNAP-Tag.

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