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在MFS糖运输机中,用于Na+合的支持器的移动屏障机制
Parameswaran Hariharan1, Yuqi Shi2, Satoshi Katsube1
1Department of Cell Physiology and Molecular Biophysics, Center for Membrane Protein Research, Texas Tech University Health Sciences Center, School of Medicine, Lubbock, TX 79424, USA.
bioRxiv : the preprint server for biology
|October 4, 2023
概括
研究人员发现了细菌的梅利菌转运体 (MelB) 如何在细胞内释放糖. 一个新的纳米体揭示了低亲和度向内面的结构,解释了基质释放和积累机制.
科学领域:
- 结构生物学 结构生物学
- 生物化学 生物化学
- 分子运输分子的运输.
背景情况:
- 阴离子结合的转运体对于细胞吸收至关重要,但它们的确切机制尚不清楚.
- 细菌的水转运体 (MelB) 作为Na+结合的主要促进体超级家族 (MFS) 转运体的模型.
研究的目的:
- 阐明糖运输和监管在MelB.的机制细节.
- 为了确定基质释放和积累的结构基础.
主要方法:
- 低温电子显微镜 (cryo-EM) 用一个构造性纳米体来获得MelB的低糖亲和度,面向内,Na+结合的结构.
- 异热定位热量计 (ITC) 用于结合试验.
- 分子动力学 (MD) 模拟.分子动力学 (MD) 模拟.
- /交换质谱 (HDX-MS) 用于识别动态区域.
主要成果:
- 通过结合面向内部和面向外的结构,定位了MelB的外部和内部屏障.
- 已识别的N端和C端残留物形成了内部屏障内的糖选择性口袋.
- 证明面向内部的形状表现出显著下降的糖结合亲和力,促进细胞内释放.
- 显示屏障转移会影响糖亲和力,但不会影响阴离子结合,得到了MD模拟的支持.
- 确定了涉及内部屏障盐桥网络的动态区域,对交通至关重要.
结论:
- 这项研究提供了关于阴阳合同承载的移动屏障机制的结构和动态见解.
- 这些发现解释了MelB如何调节糖结合亲和力,并促进基质释放和积累.
- 形状纳米体是研究传送器动态和机制的宝贵工具.
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