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4D-diaXLMS:用于交叉链接质谱的全蛋白质四维数据独立获取工作流

Yanhong Hao¹, Moran Chen¹, Xiao Huang¹

¹The Institute for Advanced Studies, Wuhan University, Wuhan, Hubei 430072, China.

Analytical chemistry

|September 11, 2023

概括

新的4D-diaXLMS方法在质谱学中显著改善了交叉链接的识别. 这种先进的技术为复杂的蛋白质组分析提供了更高的覆盖范围,可重复性和准确的量化.

科学领域:

蛋白质组学是指蛋白质组学.
生物化学生物化学
分析化学分析化学

背景情况:

交联质谱 (XL-MS) 对于蛋白质结构和相互作用研究至关重要.
数据依赖获取 (DDA) 模式在分析低丰度交联上存在局限性,因为它具有随机性.

研究的目的:

引入和评估4D-diaXLMS,这是一种使用四维数据独立获取进行全蛋白质组跨链分析的新工作流.
证明4D-diaXLMS在复杂生物样本中识别和量化交联的优势.

主要方法:

开发和应用4D-diaXLMS工作流程,用于交叉链接质谱.
使用HeLa细胞蛋白质组和disuccinimidyl suberate交叉链接器对4D-diaXLMS和DDA进行比较分析.
在复杂的样本矩阵中评估识别覆盖范围,可重现性和定量准确性.

主要成果:

4D-diaXLMS在一次性分析中实现了与DDA相比,交叉链接鉴定覆盖率的36%增加,在减少分离时,提高了高达149%.
可复制性显著提高,使用4D-diaXLMS识别了83%的交联,而使用DDA的38%是4D-diaXLMS.
该方法证明了在复杂的HeLa细胞基质中酵母交联的高定量准确性和低变化系数.

结论:

4D-diaXLMS为复杂蛋白质组中的深入XL-MS分析提供了卓越的识别覆盖,可重复性和定量准确性.
这一工作流代表了对未来的蛋白质组和结构生物学研究具有巨大潜力的重大进步.

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