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在ATP结合和基质结合状态中被困的人类ABCG2突变的冷EM结构
Ioannis Manolaridis1, Scott M Jackson1, Nicholas M I Taylor2,3
1Institute of Molecular Biology and Biophysics, Department of Biology, ETH Zurich, Switzerland.
Nature
|November 9, 2018
概括
这项研究揭示了ABCG2传送器功能的结构机制. 高分辨率的冷EM结构显示了ATP结合如何驱动细胞膜的基质转移.
科学领域:
- 生物化学
- 结构生物学
- 分子生物学
背景情况:
- ABCG2 (ATP结合盒子子子家族G成员2) 是一个关键的载体蛋白,参与细胞防御和药物药理动力学.
- 虽然ABCG2的结构和抑制已知,但其基质识别和传输机制尚不清楚.
研究的目的:
- 阐明人类ABCG2基质识别和ATP驱动的运输的分子机制.
- 在关键功能状态下呈现ABCG2的高分辨率冷电子显微镜 (cryo-EM) 结构.
主要方法:
- 使用高分辨率冷电子显微镜 (cryo-EM) 来确定功能性ABCG2突变的结构 (ABCG2EQ).
- 用以捕捉转移前和转移后状态的模型基质是雌激素-3-硫酸盐 (E1S).
- 为了验证结构发现,进行了突变酶和体外输送和ATPase测试.
主要成果:
- 化EM结构显示了ABCG2的不同基质结合 (转移前) 和ATP结合 (转移后) 状态.
- 在转移前的状态下,基质结合腔是中心的,面向细胞质,容纳单个E1S分子.
- ATP 结合会诱导形状变化,使内腔崩并为基质挤出而开辟外腔,涉及域移位和改变 NBD 方向.
结论:
- ABCG2利用ATP结合来驱动基质转移的结构变化.
- 特定的残留物,包括白"插头",对于基质的识别和基质与抑制剂的区别至关重要.
- 这些发现为ABCG2如何作为排泄发挥作用提供了机械洞察力,从而影响了药物的疗效和异生菌保护.
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