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综合蛋白质可溶性转移试验用于全面的药物标识,在全蛋白质组范围的规模

Xiaolei Zhang¹, Chengfei Ruan^1,2, Yan Wang¹

¹CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R & A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China.

Analytical chemistry

|September 7, 2023

概括

一种新的iso-pH转移测定 (ipHSA) 方法通过测量连接体诱导的蛋白质溶解度转移来增强蛋白质组范围的药物标识. 综合方法 (IPSSA) 进一步提高了灵敏度和吞吐量,为行动研究的机制提供了一个具有成本效益的工具.

科学领域:

生物化学生物化学
蛋白质组学是指蛋白质组学.
化学生物学化学生物学

背景情况:

带结合往往稳定了目标蛋白,增加了对变质的抵抗力.
目前用于识别带标蛋白的方法可能昂贵,并且具有有限的吞吐量.
需要高效,高通量方法来实现全蛋白质组范围的标识.

研究的目的:

开发一种基于连接体诱导的蛋白质可溶性转移的新型,高通量蛋白质组范围的标识方法.
改进现有的方法,如pH-依赖蛋白沉 (pHDPP).
提高药物目标发现的敏感性和降低成本.

主要方法:

开发使用数据独立获取 (DIA) 来量化蛋白质的iso-pH转移试验 (ipHSA).
将ipHSA与现有方法进行比较,例如改进的同热转移试验 (iTSA) 和同溶剂转移试验 (iSSA).
通过结合ipHSA,iTSA和iSSA的结果,创建一个综合蛋白质溶解度转移试验 (IPSSA).

主要成果:

与pHDPP相比,ipHSA在稳体菌素标识方面表现出更高的吞吐量.
在使用DIA进行量化时,ipHSA与iTSA和iSSA显示出高度的互补性.
与单个方法相比,IPSSA在选稳氨酸标时显著提高了灵敏度 (38-38%).
IPSSA提高了吞吐量和降低了成本,增加了复制品,进一步提高了灵敏度.

结论:

异位-pH转移试验 (ipHSA) 是一种有效的蛋白质组范围目标鉴定方法.
综合蛋白质可溶性转移试验 (IPSSA) 提供了增强的灵敏度,吞吐量和成本效益.
IPSSA是识别药物点和研究作用机制的宝贵工具.

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