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通过SuFEx点击化学,通过蛋白质和蛋白质复合体进行强大的化学交联

Xiaoya Li¹, Kai Li¹, Xuechun Yuan¹

¹Department of Medicinal Chemistry, School of Pharmacy, China Pharmaceutical University, Nanjing 211198, P. R. China.

Analytical chemistry

|July 8, 2025

概括

新的硫尼尔化物 (SF) 交叉连接器,利用硫化物交换 (SuFEx) 点击化学,增强蛋白质结构分析. 这些试剂具有强大的反应性,并使蛋白质和复合物的精确3D建模成为可能.

科学领域:

生物化学生物化学
化学生物学化学生物学
结构生物学结构生物学

背景情况:

蛋白质的化学交叉链接与质谱学 (CXMS) 相结合,对于了解蛋白质结构,相互作用和动态至关重要.
现有的CXMS方法需要具有增强反应性和性能的新型交叉链接器,以改进蛋白质结构建模.
扩大交叉链接覆盖范围和更高的数据质量对于推动CXMS在生物研究和疾病理解中的应用至关重要.

研究的目的:

为CXMS应用开发基于硫-化物交换 (SuFEx) 的新型同位体交叉链接器,点击化学.
调查这些新型含硫尼尔化物 (SF) 交叉连接器的反应性和效用.
为了证明SuFEx的潜力,点击化学在扩大CXMS工具箱的蛋白质结构阐明.

主要方法:

使用SuFEx点击化学合成含有硫尼尔化物 (SF) 的同位体交叉连接剂.
用SF交叉链接器来模拟,蛋白质和细胞溶解物的应用.
质谱分析以识别交联和确定蛋白质结构.
为蛋白质和蛋白质复合体生成3D结构模型.

主要成果:

开发的SF交叉连接器对氨酸和氨酸残留物表现出强大的反应性.
SF交叉链路显示出与已知的现有蛋白质结构的良好对齐.
该方法可以为缺乏先前实验结构数据的蛋白质和蛋白质复合体生成新的3D模型.

结论:

硫交换 (SuFEx) 点击化学为开发先进的化学交叉链接器提供了一个新的平台.
SF交叉链接器显著扩大了CXMS用于蛋白质结构确定的能力.
这种方法提供了与当前交叉连接试剂无法获得的补充结构信息,推进了结构生物学领域.

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