Induced degradation of SNAP-fusion proteins†

IF 4.2 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

RSC Chemical Biology Pub Date : 2024-10-21 DOI:10.1039/D4CB00184B

Savina Abraham Pol, Sara Liljenberg, Jack Barr, Gina Simon, Luis Wong-Dilworth, Danielle L. Paterson, Vladimir P. Berishvili, Francesca Bottanelli, Farnusch Kaschani, Markus Kaiser, Mariell Pettersson and Doris Hellerschmied

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Abstract

Self-labeling protein tags are an efficient means to visualize, manipulate, and isolate engineered fusion proteins with suitable chemical probes. The SNAP-tag, which covalently conjugates to benzyl–guanine and –chloropyrimidine derivatives is used extensively in fluorescence microscopy, given the availability of suitable SNAP-ligand-based probes. Here, we extend the applicability of the SNAP-tag to targeted protein degradation. We developed a set of SNAP PROteolysis TArgeting Chimeras (SNAP-PROTACs), which recruit the VHL or CRBN-ubiquitin E3 ligases to induce the degradation of SNAP-fusion proteins. Endogenous tagging enabled the visualization and the selective depletion of a SNAP-clathrin light chain fusion protein using SNAP-PROTACs. The addition of PROTACs to the SNAP-tag reagent toolbox facilitates the comprehensive analysis of protein function with a single gene tagging event.

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诱导 SNAP 融合蛋白降解。

自标记蛋白质标签是利用合适的化学探针观察、操作和分离工程融合蛋白的有效方法。SNAP 标签能与苄基鸟嘌呤和氯嘧啶衍生物共价结合，由于有合适的基于 SNAP 配体的探针，它在荧光显微镜中得到了广泛应用。在这里，我们将 SNAP 标记的适用性扩展到靶向蛋白质降解。我们开发了一组 SNAP PROteolysis TArgeting Chimeras（SNAP-PROTACs），它们能招募 VHL 或 CRBN-ubiquitin E3 连接酶来诱导 SNAP 融合蛋白的降解。使用 SNAP-PROTACs 对 SNAP-clathrin 轻链融合蛋白进行内源标记可实现可视化和选择性降解。在 SNAP 标记试剂工具箱中加入 PROTACs 后，只需一次基因标记就能对蛋白质功能进行全面分析。

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来源期刊

RSC Chemical Biology Multiple-

CiteScore

6.10

自引率

0.00%

发文量

128

审稿时长

10 weeks