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超微小的化学遗传标签与组转移配体

Sreekanth Vedagopuram^1,2, Shaimaa Sindi^1,2, Santosh K Chaudhary^1,2

¹Chemical Biology and Therapeutics Science, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.

bioRxiv : the preprint server for biology

|July 14, 2025

概括

研究人员为蛋白质研究开发了超小化学遗传标签 (mgTag和cTag). 这些新标签是报告中最小的标签,可以在不破坏功能的情况下精确修改蛋白质,从而推动生物技术和医学的发展.

科学领域:

生物化学生物化学
分子生物学分子生物学
化学生物学化学生物学

背景情况:

化学遗传标签对于研究蛋白质功能至关重要,当小分子配体无法使用时.
现有的标签往往太大,限制了它们的应用与某些感兴趣的蛋白质 (POI).

研究的目的:

开发和描述用于蛋白质研究的新型,超微小的化学遗传标签.
为了证明这些标签在细胞信号和近距离诱导应用中的实用性.

主要方法:

设计了两种超小化学遗传标签,mgTag (36 aa) 和cTag (50 aa),具有转移酶类型的反应性.
在标签内利用工程氨酸残留物,用于用特定带进行组转移反应.
通过将它们与KRASG12D和阿贝尔森酶 (ABL) 融合,并通过诱导ABL和BRD4之间的接近来评估标签的功能.

主要成果:

据报道,mgTag和cTag是最小的化学遗传标签,使得功能蛋白质的研究能够不受干扰.
与较大的HaloTag不同,mgTag或cTag与KRASG12D的融合不会影响其生长信号通路的激活.
使用mgTag/cTag和BRD4结合剂实现了ABL和BRD4之间的近距离诱导,从而导致BRD4酸化.

结论:

超微化学遗传标签 (mgTag,cTag) 为研究蛋白质功能和工程生物系统提供了重大进展.
这些标签的转移酶类型反应性和小尺寸为基础科学,生物技术和医学提供了多方面的应用.

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Tagging and Fusion Proteins

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Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...