在MEK-RAF/KSR-14-3-3复合体上化MEKi的活细胞向激活
William M Marsiglia1,2,3, Arthur Chow4,5,6, Zaigham M Khan4,5
1Department of Oncological Sciences, The Tisch Cancer Institute, Mount Sinai Center for Therapeutic Discovery, Icahn School of Medicine at Mount Sinai, New York, NY, USA. wmmarsig@uab.edu.
Nature chemical biology
|November 3, 2023
概括
新的NanoBRET测试测量了活细胞中针对性药物与RAS-MAPK通路复合物的接触. 这些测试揭示了不同的抑制剂结合概况,并可以选特定途径标的化合物.
科学领域:
- 分子生物学分子生物学
- 蜂信号传输是如何进行的
- 药理学 药理学是指药理学的学科.
背景情况:
- RAS-mitogen激活蛋白激酶 (MAPK) 途径在细胞信号和疾病中至关重要.
- 向疗法,如全性MEK抑制剂 (MEKi),显示出希望,但它们的有效性取决于与特定途径复合物的相互作用.
- 在活细胞中直接测量这些生理复合体内的药物相互作用是一个重大挑战.
研究的目的:
- 开发和验证基于NanoBRET的测试,以量化MEK抑制剂 (MEKi) 与MEK1及其相关复合物的活细胞中的接触.
- 描述不同RAS-MAPK通路组件的MEKi结合配置文件,包括ARAF,BRAF,CRAF,KSR1和KSR2,有或没有14-3-3.
- 评估病原性突变,如BRAF-V600E,对MEKi结合的影响.
主要方法:
- 开发NanoBRET测试以测量MEKi的直接目标参与度.
- 在活细胞中量化MEKi与MEK1结合复合物的相互作用,其中包括ARAF,BRAF,CRAF,KSR1和KSR2.
- 包括14-3-3蛋白来模仿生理条件和评估BRAF-V600E突变的影响.
主要成果:
- 在各种MEK1结合的复合体中观察到明显的MEKi偏好.
- 基于这些观察到的偏好,生成了抑制剂结合概况.
- 这些分析成功报告了BRAF-V600E突变对MEKi结合的影响.
结论:
- 开发的NanoBRET试验为测量MEKi在活细胞中的目标参与提供了一个强大的平台.
- 这些测试可以生成RAS-MAPK通路复合体的特定抑制剂结合概况.
- 该平台可用于选用于治疗开发的RAS-MAPK级联中的特定复合体的化合物.
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