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用于肌动蛋白可视化的新型香豆素荧光染料的合成。

Synthesis of First Coumarin Fluorescent Dye for Actin Visualization.

机构信息

School of Chemistry, Sambalpur University, Jyoti Vihar, Burla, Sambalpur 768019, Odisha, India.

Neural Developmental Biology Lab, Department of Life Science, National Institute of Technology, Rourkela 769008, Odisha, India.

出版信息

Bioconjug Chem. 2022 Nov 16;33(11):2113-2120. doi: 10.1021/acs.bioconjchem.2c00332. Epub 2022 Oct 20.

Abstract

Selective fluorescence imaging of actin protein hugely depends on the fluorescently labeled actin-binding domain (ABD). Thus, it is always a challenging task to image the actin protein ( or ) directly with an ABD-free system. To overcome the limitations of actin imaging without an ABD, we have designed a facile and cost-effective red fluorescent coumarin dye 7-hydroxy-4-methyl-8-(4-(2-oxo-2-chromen-3-yl)thiazol-2-ylimino)methyl-2-chromen-2-one () for actin binding. The selective binding of the dye was investigated using the gut and eye of the model organism and and cell lines. Our result suggests two major advantages of over the dyes presently used for imaging actin proteins. First, the dye can bind to actin efficiently without any secondary intermediate. Second, it is much more stable at room temperature and exhibits excellent photostability. To the best of our knowledge, this is the first fluorescent dye that can bind to the actin protein without employing any secondary intermediate/actin-binding domain. These findings could pave the way for many biologists and physicists to successfully employ the dye for imaging and tracking actin proteins by fluorescence microscopy in various and systems.

摘要

肌动蛋白蛋白的选择性荧光成像在很大程度上取决于荧光标记的肌动蛋白结合域 (ABD)。因此,直接用无 ABD 的系统来成像肌动蛋白蛋白 (或) 一直是一项具有挑战性的任务。为了克服无 ABD 成像的局限性,我们设计了一种简单且具有成本效益的红色荧光香豆素染料 7-羟基-4-甲基-8-(4-(2-氧代-2-色烯-3-基)噻唑-2-基亚氨基)甲基-2-色烯-2-酮 () 用于肌动蛋白结合。使用模型生物 和 以及 和 细胞系研究了染料的选择性结合。我们的结果表明,与目前用于成像肌动蛋白蛋白的染料相比, 具有两个主要优势。首先,该染料可以在没有任何二级中间体的情况下有效地与肌动蛋白结合。其次,它在室温下更稳定,表现出优异的光稳定性。据我们所知,这是第一个可以在不使用任何二级中间体/肌动蛋白结合域的情况下与肌动蛋白蛋白结合的荧光染料。这些发现可能为许多生物学家和物理学家铺平道路,使他们能够成功地在各种 和 系统中通过荧光显微镜使用 染料对肌动蛋白蛋白进行成像和追踪。

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