Photo-SNAP-tag, a Light-Regulated Chemical Labeling System.,ACS Chemical Biology

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Photo-SNAP-tag, a Light-Regulated Chemical Labeling System.
ACS Chemical Biology ( IF 3.5 ) Pub Date : 2020-07-05 , DOI: 10.1021/acschembio.0c00412
Joseph D Cleveland ₁ , Chandra L Tucker ₁

Affiliation

Methods that allow labeling and tracking of proteins have been instrumental for understanding their function. Traditional methods for labeling proteins include fusion to fluorescent proteins or self-labeling chemical tagging systems such as SNAP-tag or Halo-tag. These latter approaches allow bright fluorophores or other chemical moieties to be attached to a protein of interest through a small fusion tag. In this work, we sought to improve the versatility of self-labeling chemical-tagging systems by regulating their activity with light. We used light-inducible dimerizers to reconstitute a split SNAP-tag (modified human O⁶-alkylguanine-DNA-alkyltransferase, hAGT) protein, allowing tight light-dependent control of chemical labeling. In addition, we generated a small split SNAP-tag fragment that can efficiently self-assemble with its complement fragment, allowing high labeling efficacy with a small tag. We envision these tools will extend the versatility and utility of the SNAP-tag chemical system for protein labeling applications.

中文翻译：

Photo-SNAP-tag，一种光调节化学标记系统。

允许标记和跟踪蛋白质的方法有助于理解它们的功能。标记蛋白质的传统方法包括与荧光蛋白融合或自标记化学标记系统，如 SNAP-tag 或 Halo-tag。这些后一种方法允许通过小的融合标签将明亮的荧光团或其他化学部分连接到感兴趣的蛋白质上。在这项工作中，我们试图通过用光调节其活动来提高自标记化学标记系统的多功能性。我们使用光诱导二聚体来重建分裂的 SNAP 标签（修饰的人类 O ⁶-烷基鸟嘌呤-DNA-烷基转移酶，hAGT）蛋白质，允许对化学标记进行严格的光依赖控制。此外，我们生成了一个小的分裂 SNAP 标签片段，它可以有效地与其补体片段自组装，从而使用小标签实现高标记效率。我们设想这些工具将扩展 SNAP-tag 化学系统在蛋白质标记应用中的多功能性和实用性。

更新日期：2020-08-21

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