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通过合法实现一个35-Plex 串联质量标记反应剂通过合法设置

Nathan R Zuniga¹, Dustin C Frost², Karsten Kuhn³

¹Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, United States.

Journal of proteome research

|October 9, 2024

概括

我们使用提升了TMTpro试剂多重复合到35-plex. 一种新的策略 (DISAT) 纠正效应,使细胞系和化学蛋白质组学中精确的蛋白质量定量成为可能.

科学领域:

蛋白质组学是指蛋白质组学.
分析化学分析化学
生物化学生物化学

背景情况:

基于质谱的样本多重复合与异标签使得高通量定量生物测试.
TMTpro试剂组允许进行多重蛋白质组分析.
将纳入等离子标签可以增加复杂化,但可能会影响定量准确性.

研究的目的:

为了将TMTpro试剂的多重复合能力几乎翻一番,设置为35-plex.
制定一种策略,以抵消由合体所引起的低于最佳峰值凝.
在生物和化学蛋白质组学实验中验证新的复合策略.

主要方法:

将一个同位素纳入TMTpro试剂的报告组.
制定"设计独立的子综合体,但一起获得" (DISAT) 规范化战略.
将非和含的道分隔成不同的子集,并通过共同的桥梁道重新组装.
在人类细胞系的比较蛋白质组学和针对Pin1的化学蛋白质组学实验中应用DISAT策略.

主要成果:

实现了TMTpro试剂的35倍复合能力.
DISAT战略有效地弥补了引起的量化错误.
在比较人类细胞系之间的蛋白质表达差异方面证明了该方法的成功应用.
使用开发的化学蛋白质组学方法,确定了与Pin1的氨酸-113结合的化合物.

结论:

修改后的TMTpro试剂组和DISAT策略显著提高了基于质谱的蛋白质组学的多重复合能力.
这种方法在定量测量中保持了很高的准确性,即使加入了.
经过验证的方法为高通量全蛋白质组研究和向化学蛋白质组学提供了强大的工具.

关键词:

这是ABPPPP ABPPPP.星际天文星际天文 TMTpro TMTpro 是一个专业的项目.TMTproD 的使用方法德是最重要的元素.异酸标记是同酸标记的

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