Activation of Nanoscale Allosteric Protein Domain Motion Revealed by Neutron Spin Echo Spectroscopy
Activation of Nanoscale Allosteric Protein Domain Motion Revealed by Neutron Spin Echo Spectroscopy
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DOI：
10.1016/j.bpj.2010.09.058
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发表时间：
2010-11-17
影响因子：
3.4
通讯作者：
Bu, Zimei
中科院分区：
文献类型：
--
作者：
Farago, Bela;Li, Jianquan;Bu, Zimei
NHERF1是一种多结构域支架蛋白，可组装信号复合物，调节多种膜蛋白的细胞表面表达和内吞循环。NHERF1中两个PDZ结构域组装蛋白复合物的能力受到膜-细胞骨架连接蛋白ezrin的变弹性调节，其结合位点位于距离PDZ结构域110埃的地方。本文利用中子自旋回波（NSE）光谱、选择性氘标记和理论分析，揭示了NHERF1在与ezrin形成配合物时在纳米长度尺度和亚微秒时间尺度上的畴间运动激活。我们表明，一个非常简化的粗粒度模型足以描述多结构域蛋白质或蛋白质复合物的结构域间运动。我们期望未来的NSE实验将受益于利用我们的选择性氘化方法，从大量的全球平移和旋转运动中解决感兴趣的特定区域运动。我们的研究结果表明，变构信号向远端位点的动态传播涉及亚微秒时间尺度上的远程耦合域运动的变化，并且这些耦合运动可以用NSE来区分和表征。
NHERF1 is a multidomain scaffolding protein that assembles signaling complexes, and regulates the cell surface expression and endocytic recycling of a variety of membrane proteins. The ability of the two PDZ domains in NHERF1 to assemble protein complexes is allosterically modulated by the membrane-cytoskeleton linker protein ezrin, whose binding site is located as far as 110 angstrom ngstroms away from the PDZ domains. Here, using neutron spin echo (NSE) spectroscopy, selective deuterium labeling, and theoretical analyses, we reveal the activation of interdomain motion in NHERF1 on nanometer length-scales and on submicrosecond timescales upon forming a complex with ezrin. We show that a much-simplified coarse-grained model suffices to describe interdomain motion of a multidomain protein or protein complex. We expect that future NSE experiments will benefit by exploiting our approach of selective deuteration to resolve the specific domain motions of interest from a plethora of global translational and rotational motions. Our results demonstrate that the dynamic propagation of allosteric signals to distal sites nvolves changes in long-range coupled domain motions on submicrosecond timescales, and that these coupled motions can be distinguished and characterized by NSE.