发现使用蛋白质组范围稳定性测试的RNA-蛋白质分子
Stanley I Goldstein1,2,3, Alice C Fan1,2, Zihao Wang2
1BU Target Discovery & Proteomics Laboratory (BU-TDPL), Boston University, Boston, MA, USA.
bioRxiv : the preprint server for biology
|April 25, 2024
概括
这项研究引入了一种新的化学蛋白质组学方法,以发现基质依赖性抑制剂,识别了与DEAD-box螺旋酶和RNA结合的Rocaglate药物 (如Zotatifin) 的新标.
科学领域:
- 化学生物学 化学生物学
- 蛋白质组学是指蛋白质组学
- 药物发现 药物发现 药物发现
背景情况:
- 非竞争性抑制对于酶调节至关重要,但由于结构数据有限,对联体的发现具有挑战性.
- 在蛋白质组学中依赖基质的向参与研究很少,阻碍了新型抑制剂的开发.
研究的目的:
- 开发一种发现基质依赖性连接体结合的策略.
- 为了识别新型目标的rocaglates,一种与RNA和DEAD-box螺旋酶相互作用的分子类.
主要方法:
- 蛋白质组完整溶解性改变 (PISA) 试验检测细胞溶解物中的RNA-蛋白质-小分子复合体.
- 目标类特定的热窗口和ATP模拟和RNA依赖性的分析.
- 有限的蛋白质溶解质谱和光极化 (FP) 用于目标验证和结构洞察.
主要成果:
- 使用PISA试验证明了一种检测RNA-蛋白质-小分子复合物的方法.
- 已确定并验证了新型的DEAD-box (DDX) 化酶标,包括临床候选物zototifin.
- 提供了关于rocaglate与DDX3X的结合亲缘关系的结构见解.
结论:
- 开发的化学蛋白质组学方法为选非竞争性抑制剂提供了一个模型.
- 发现了可操作的DDX紧目标,为新的分子紧和RNA螺旋酶表征铺平了道路.
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