细菌载体EmrE在脂层中的微秒运动
Alexander A Shcherbakov1, Merissa Brousseau2, Katherine A Henzler-Wildman2
1Department of Chemistry, Massachusetts Institute of Technology, 170 Albany Street, Cambridge, Massachusetts 02139, United States.
细菌输送器EmrE使用微秒的骨干运动来促进基质的结合和释放. 在与四 () 结合的突变体中观察到的这些动态,为多药耐药传递机制提供了洞察力.
科学领域:
- 生物化学
- 结构生物学
- 膜运输
背景情况:
- EmrE是一种细菌载体,也是小型多药性 (SMR) 蛋白质的模型.
- 它流出与质子运输合的阴离子基质.
- 了解EMR的结构动态关系是阐明SMR传送机制的关键.
研究的目的:
- 为了研究 EmrE 与基质复合的蛋白质动态.
- 在运输过程中将蛋白质运动与基质结合和释放相关联.
主要方法:
- 在脂质双层中使用固态NMR光谱对基质结合的S64V- EmrE突变物进行检测.
- 在魔法角旋转 (MAS) 下测量了N个旋转框架旋转放松率 (R1ρ).
- 在55kHz的MAS下进行了1个H检测的15个旋锁实验.
主要成果:
- 测量了基质结合蛋白的特定部位放松率.
- 在280K时,旋锁取决于场的放松指示了脊柱运动.
- 这些运动发生在酸性和性 pH 时,独立于质子化状态.
结论:
- 在基板绑定的EmrE中存在微秒时间尺度的骨干运动.
- 这些运动比整个运输周期更快,但与基质结合/释放时间表保持一致.
- EmrE可能利用这些动态运动来采样有效的基板运输.
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