<br>用嘌呤霉素光电开关对平移进行光学控制,Journal of the American Chemical Society

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用嘌呤霉素光电开关对平移进行光学控制
Journal of the American Chemical Society ( IF 14.4 ) Pub Date : 2022-11-17 , DOI: 10.1021/jacs.2c07374
Tongil Ko ₁ , Mauricio M Oliveira ₂ , Jessica M Alapin ₂ , Johannes Morstein ₁ , Eric Klann ₂ , Dirk Trauner ₁

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翻译是一个基本的细胞过程，涉及大量因素与核糖体以协调一致的方式相互作用。已经出现了许多干扰核糖体功能的天然产物，例如嘌呤霉素，它模仿氨酰基 tRNA 并导致链过早终止。在这里，我们介绍了嘌呤霉素的光可切换版本，它实际上将翻译置于光学控制之下。我们的化合物称为puroswitch ，具有重氮辛，可实现可逆且近乎定量的异构化和药理调节。其合成涉及一种新的光可切换氨基酸结构单元。 Puroswitch在黑暗中表现出很少的活性，并且在不同波长的可见光照射下以分级方式变得更加活性和细胞毒性。体外翻译测定证实， puroswitch通过与嘌呤霉素本身类似的机制抑制翻译。一旦掺入新生蛋白质中， puroswitch就会与标准嘌呤霉素抗体发生反应，从而可以使用蛋白质印迹和免疫组织化学来追踪蛋白质的从头合成。作为一种细胞渗透性小分子， puroswitch可用于多种细胞类型（包括原代小鼠神经元）的新生蛋白质组分析。我们设想puroswitch作为一种有用的生化工具，用于光学控制翻译和在确定的位置和精确的时间点监测新合成的蛋白质。

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Optical Control of Translation with a Puromycin Photoswitch

Translation is an elementary cellular process that involves a large number of factors interacting in a concerted fashion with the ribosome. Numerous natural products have emerged that interfere with the ribosomal function, such as puromycin, which mimics an aminoacyl tRNA and causes premature chain termination. Here, we introduce a photoswitchable version of puromycin that, in effect, puts translation under optical control. Our compound, termed puroswitch, features a diazocine that allows for reversible and nearly quantitative isomerization and pharmacological modulation. Its synthesis involves a new photoswitchable amino acid building block. Puroswitch shows little activity in the dark and becomes substantially more active and cytotoxic, in a graded fashion, upon irradiation with various wavelengths of visible light. In vitro translation assays confirm that puroswitch inhibits translation with a mechanism similar to that of puromycin itself. Once incorporated into nascent proteins, puroswitch reacts with standard puromycin antibodies, which allows for tracking de novo protein synthesis using western blots and immunohistochemistry. As a cell-permeable small molecule, puroswitch can be used for nascent proteome profiling in a variety of cell types, including primary mouse neurons. We envision puroswitch as a useful biochemical tool for the optical control of translation and for monitoring newly synthesized proteins in defined locations and at precise time points.

更新日期：2022-11-17

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