针对药物标的蛋白质体的缩小式热分析和凝聚分析以及蛋白质相互作用分析
Xue Lu1, Bin Liao1, Siyuan Sun1
1Department of Chemistry, Southern University of Science and Technology, Shenzhen 518055, China.
Analytical chemistry
|September 1, 2023
概括
我们开发了STASIS,一种缩小规模的热蛋白质组分析 (TPP) 方法. 这种具有成本效益的技术需要最小的蛋白质用于药物向解和罕见细胞中的蛋白质复合体分析.
科学领域:
- 生物化学 生化学
- 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:
- 热蛋白质组概况 (TPP) 和热近距离聚合 (TPCA) 识别药物蛋白相互作用和蛋白质复合体动态.
- 目前的TPP/TPCA方法每样需要100微克以上的蛋白质,限制其用于罕见细胞和临床样本.
研究的目的:
- 开发一个缩小的工作流程 (STASIS) 用于热蛋白质组概况和TPCA,大大降低了蛋白质输入要求.
- 评估STASIS工作流在蛋白质识别,复杂分析和药物向解方面的性能.
主要方法:
- 开发了STASIS (缩小规模的热分析和与SISPROT的凝聚分析),将CETSA与MS集成.
- 使用SISPROT技术和基于尖端的分化来处理PCR管中加热的样品.
- 评估STASIS的蛋白质输入量为每温度10,5和1μg.
主要成果:
- STASIS识别了40005000种蛋白质,采集时间为6小时,每温度使用的蛋白质仅为1μg.
- 在不同蛋白质输入中观察到化温度 (Tm) 的高相关性和可比的TPCA性能.
- 通过使用1μg的蛋白质,成功地识别了高精度的甲醇和泛基的标.
结论:
- STASIS是一种强大的,具有成本效益的技术,可使TPP和TPCA具有显著减少的蛋白质量.
- 工作流程将热蛋白质组分析和TPCA的应用扩展到罕见的原始细胞和宝贵的临床样本.
- 在低投入的生物样本中,STASIS促进了精确的药物向解和蛋白质复合体分析.
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