一个不可逆转的agonist-β(2) 上腺受体复合物的结构和功能
Daniel M Rosenbaum1, Cheng Zhang, Joseph A Lyons
1Department of Molecular and Cellular Physiology, Stanford University School of Medicine, 279 Campus Drive, Stanford, California 94305, USA.
Nature
|January 14, 2011
概括
研究人员开发了一种用于β-2上腺体受体 (β(2) AR) 的新型共价激动剂. 这使受体稳定了受体.
科学领域:
- 结构生物学是结构生物学.
- 生物化学 生物化学
- 分子药理学分子药理学
背景情况:
- G蛋白结合受体 (GPCR) 是关键的细胞信号蛋白,但激素结合机制尚不清楚.
- 了解GPCR激活是药物设计的关键,但结晶激素结合状态是由于弱联体相互作用的挑战.
- 人类β-2上腺素受体 (β(2) AR) 是一个经过充分研究的GPCR,但其活性构造很难从结构上捕获.
研究的目的:
- 为了确定一种与激动剂结合的GPCR的高分辨率结构.
- 阐明GPCR激活和全调节的分子基础.
- 为了使新型GPCR向治疗的结构导向设计.
主要方法:
- 设计和合成一种新型的对抗β(2) AR的共价激活剂,通过二硫化键连接.
- 形成一个能够激活G蛋白的稳定共价β(2) AR-agonist复合体.
- 在脂质双层中结晶共价激剂结合的β(2) AR-T4L融合蛋白,使用脂质中位相法.
- 在3.5 Å分辨率下确定复合物的结构.
主要成果:
- 一个稳定的共价激动剂结合的β(2) AR复合物成功地被制造和结晶.
- 结构分析揭示了受体的活性构造,需要在细胞外和细胞内表面相互作用.
- 分子动力学模拟 (高达30微秒) 显示,在没有G蛋白或稳定抗体的情况下,活性状态的不稳定性.
结论:
- 一种共性激动剂策略克服了结晶激动剂结合的GPCRs方面的挑战.
- 确定的结构为β(2) AR激活所需的形状变化提供了关键的见解.
- 这项工作促进了对GPCR激活机制的理解,并促进了基于结构的药物发现.
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