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低背景His-Tag准探针用于启动细胞表面蛋白质及其结合相互作用的光检测

Pragati Kishore Prasad¹, Suraj Toraskar¹, Suman Khan²

¹Department of Chemical and Structural Biology, Weizman Institute of Science, Rehovot, 7610001, Israel.

Small (Weinheim an der Bergstrasse, Germany)

|July 4, 2025

概括

研究人员开发了新的光探针来检测细胞表面蛋白质及其相互作用. 这种新方法简化了针对SARS-CoV-2病毒受体结合域 (RBD) 和ACE2相互作用的抑制剂的测试.

科学领域:

生物化学生物化学
化学生物学化学生物学
分子成像学分子成像学

背景情况:

开启式光探针对于研究细胞表面蛋白 (CSP) 和它们的相互作用至关重要.
开发基于蛋白质的配体探针具有挑战性,因为需要COVID-19研究试验.
在SARS-CoV-2受体结合域 (RBD) 和ACE2受体之间的相互作用需要有效的抑制剂评估.

研究的目的:

创建一个新的开启式光探针系统,用于检测CSP及其相互作用.
开发一种基于细胞的简化试验,用于检测RBD-ACE2相互作用的抑制剂.
为了证明探头系统在检测其他细胞表面事件方面的多功能性.

主要方法:

将三二三酸 (tri-NTA) 单位结合为基于素的氨酸 (QBC) 染料.
利用tri-NTA对His-tag的高亲和力和QBC染料的光学特性.
开发一种基于细胞的测试方法,将启动探针,His标记的RBD和ACE2表达细胞结合起来.

主要成果:

三-NTA-QBC探针在细胞表面与His标记蛋白结合时表现出光.
建立了一个简单的基于细胞的测定方法,通过监测光谱变化来检测RBD-ACE2相互作用的抑制剂.
这些探头成功地被用于检测细胞表面甘氨酸与莱克结合的检测,并在无洗条件下对细菌CSP进行成像.

结论:

开发的开启式光探针为研究CSP及其相互作用提供了一种强大而简单的方法.
这种方法为药物发现提供了有价值的工具,特别是病毒抑制剂.
该探针系统在基于细胞的测试和生物成像中展示了广泛的应用性.

关键词:

他的标签绑定探头.蛋白质表面识别蛋白质表面识别受体 - 连接体相互作用.提亚醇色是一种开启光探针的光探针.

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