Four-colour FRET reveals directionality in the Hsp90 multicomponent machinery
Four-colour FRET reveals directionality in the Hsp90 multicomponent machinery
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DOI：
10.1038/ncomms5192
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发表时间：
2014-06-01
影响因子：
16.6
通讯作者：
Hugel, T.
中科院分区：
文献类型：
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作者：
Ratzke, C.;Hellenkamp, B.;Hugel, T.
在生物体中，大多数蛋白质以复合物的形式工作，形成多组分蛋白质机器。这种多部件机器的功能通常是通过将它们划分为两个平衡状态系统的集合来解决的。许多分子机器，如Hsp90，通过利用ATP水解的能量而远离平衡。在这些情况下，重要的信息是通过观察连续两个以上状态的连续获得的。我们开发了一种四色单分子FRET系统来观察热休克蛋白90 （Hsp90）系统的状态序列，该系统由Hsp90二聚体、cochaperone p23和核苷酸组成。我们证明了这个多组分系统是一个定向的atp依赖机制。这揭示了先前描述的关于协伴侣蛋白如何修饰Hsp90的机制，即通过加强ATP水解与Hsp90系统中涉及的动力学步骤之间的耦合，从而产生更强的方向性。
In living organisms, most proteins work in complexes to form multicomponent protein machines. The function of such multicomponent machines is usually addressed by dividing them into a collection of two state systems at equilibrium. Many molecular machines, like Hsp90, work far from equilibrium by utilizing the energy of ATP hydrolysis. In these cases, important information is gained from the observation of the succession of more than two states in a row. We developed a four-colour single-molecule FRET system to observe the succession of states in the heat shock protein 90 (Hsp90) system, consisting of an Hsp90 dimer, the cochaperone p23 and nucleotides. We show that this multicomponent system is a directional ATP-dependent machinery. This reveals a previously undescribed mechanism on how cochaperones can modify Hsp90, namely by strengthening of the coupling between ATP hydrolysis and a kinetic step involved in the Hsp90 system resulting in a stronger directionality.