The interaction of Hsp104 with yeast prion Sup35 as analyzed by fluorescence cross-correlation spectroscopy.oligomeric state.
The interaction of Hsp104 with yeast prion Sup35 as analyzed by fluorescence cross-correlation spectroscopy.oligomeric state.
复制标题
通过荧光互相关光谱分析 Hsp104 与酵母朊病毒 Sup35 的相互作用。寡聚状态。
DOI：
10.1016/j.bbrc.2013.10.147
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发表时间：
2013
期刊：
影响因子：
--
通讯作者：
Taguchi H.
中科院分区：
文献类型：
--
作者：
Ohta S.;Kawai-Noma S.;Kitamura A.;Pack C.G.;Kinjo M.;Taguchi H.
朊病毒是一种自我繁殖的淀粉样蛋白。酵母朊病毒[PSI+]是一种基于蛋白质的可遗传元件，其中Sup 35蛋白的淀粉样蛋白聚集体被传递到子细胞。Hsp 104是一种ATP依赖性解聚酶，它和其他分子伴侣对维持[PSI+]是必不可少的。虽然先前的报道已经证实了Hsp 104和Sup 35淀粉样蛋白的物理相互作用，但Hsp 104如何与Sup 35淀粉样蛋白相互作用的机制仍有待阐明。在这里，我们研究了热休克蛋白104和Sup 35之间的相互作用[PSI+]细胞裂解液中使用荧光互相关光谱（FCCS），它可以分析不同的荧光团的共扩散事件。FCCS分析表明，在[PSI+]裂解物中，Hsp 104和Sup 35之间存在强烈的相互作用，但在[psi-]裂解物中则没有，这表明Hsp 104识别Sup 35的淀粉样蛋白聚集体。虽然保留在ATP耗尽的[PSI+]裂解物的相互作用，除了ATP或盐酸胍，这是一种抑制剂的Hsp 104，[PSI+]裂解物削弱的相互作用。
Prions are self-propagating amyloids. Yeast prion [PSI+] is a protein-based heritable element, in which amyloid aggregates of the Sup35 protein are transmitted to daughter cells. Hsp104, an ATP-dependent disaggregase, and other chaperones are essential to maintain [PSI+]. Although previous reports have demonstrated the physical interactions of Hsp104 and Sup35 amyloids, the mechanism how Hsp104 interacts with Sup35 amyloids remains to be elucidated. Here we investigated the interaction between Hsp104 and Sup35 in the lysates of [PSI+] cells using fluorescence cross-correlation spectroscopy (FCCS), which can analyze the codiffusion events of different fluorophores. FCCS analysis showed a strong interaction between Hsp104 and Sup35 in [PSI+] lysates, but not in [psi−] lysates, suggesting that Hsp104 recognizes the amyloid aggregates of Sup35. Although the interaction was retained in ATP-depleted [PSI+] lysates, addition of ATP or guanidine hydrochloride, which is an inhibitor of Hsp104, to [PSI+] lysates weakened the interaction.