基团转移配体的超小化学发生标签。

IF 16.9
Vedagopuram Sreekanth, Shaimaa H Sindi, Santosh K Chaudhary, Rajaiah Pergu, Prashant Singh, Endri Karaj, Surached Siriwongsup, Jeffrey E Fung, Arghya Deb, Stephan J DeCarlo, Jaron A M Mercer, Kei Yamada, Diego Rodriguez, David R Liu, Amit Choudhary
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引用次数: 0

摘要

化学发生标签有助于探索缺乏小分子配体的感兴趣蛋白(POI)的活性;然而,对于几个poi来说,大多数标签都太大了。在这里,我们报道了两个超小化学发生标签(mgTag和cTag),分别有36和50个氨基酸(aa),据我们所知,它们是最小的。这些标签与其配体表现出转移酶类型的反应性,允许将任何感兴趣的部分附着到标签上。cTag利用一种工程的含C1结构域的半胱氨酸与其配体发生基团转移反应。同样,mgTag利用工程锌指结构域半胱氨酸,在小脑(CRBN)存在下与其分子胶配体发生基团转移反应。虽然HaloTag (297 aa)或SNAPTag (182 aa)与KRASG12D (188 aa)的融合破坏了其生长信号通路,但mgTag或cTag的融合并未破坏其生长信号通路,这表明标签大小的重要性。BRD4结合物基团转移到Abelson激酶(ABL)上的标签上,诱导ABL和BRD4接近,导致后者磷酸化。转移酶型反应性的缺失降低了磷酸化水平,这表明具有基团转移设计的邻近诱导嵌合体可能更有效。我们设想这些超小型标签将有广泛的应用,包括基础科学、生物技术和医学。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Ultrasmall Chemogenetic Tags with Group-Transfer Ligands.

Chemogenetic tags facilitate exploration of activities of a protein-of-interest (POI) that lacks small-molecule ligands; however, most tags are too large for several POIs. Here, we report two ultrasmall chemogenetic tags (mgTag and cTag) of 36 and 50 amino acids (aa) that, to the best of our knowledge, are the smallest. These tags exhibit transferase-type reactivity with their ligands, allowing the attachment of any moiety-of-interest to the tag. cTag utilizes an engineered C1 domain-bearing cysteine that undergoes a group-transfer reaction with its ligand. Likewise, mgTag utilizes an engineered zinc-finger domain-bearing cysteine that undergoes a group-transfer reaction with its molecular glue ligand in the presence of cereblon (CRBN). While the fusion of HaloTag (297 aa) or SNAPTag (182 aa) to the KRASG12D (188 aa) disrupted its growth-signaling pathway, fusion of mgTag or cTag did not, pointing to the importance of tag size. Group-transfer of BRD4 binder to tags appended to Abelson kinase (ABL) induced proximity between ABL and BRD4, resulting in the latter's phosphorylation. Deletion of the transferase-type reactivity reduced phosphorylation levels, suggesting that proximity-inducing chimeras with group-transfer design may be more efficacious. We envision these ultrasmall tags to have wide-ranging applications, including in basic science, biotechnology, and medicine.

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