Productive interaction of chaperones with substrate protein domains allows correct folding of the downstream GFP domain.
Productive interaction of chaperones with substrate protein domains allows correct folding of the downstream GFP domain.
复制标题
伴侣与底物蛋白结构域的有效相互作用允许下游 GFP 结构域的正确折叠。
DOI：
10.1016/j.gene.2005.01.019
复制
发表时间：
2005
期刊：
影响因子：
--
通讯作者：
Chong,Shaorong
中科院分区：
文献类型：
--
作者：
Zhang,Aihua;Cantor,EricJ;Barshevsky,Tanya;Chong,Shaorong
绿色荧光蛋白(GFP)通过将溶解度与荧光相关联来报告蛋白质折叠。在GFP融合蛋白中，上游易于聚集的结构域可以扰乱大肠杆菌中GFP结构域的从头折叠，导致荧光损失。在此之前，我们证明了在蛋白质合成过程中，通过耦合的折叠和结合作用来防止上游容易聚集的结构域的错误折叠，恢复了GFP的荧光和溶解性。由于分子伴侣经常通过结合和释放相互作用折叠新生多肽，问题仍然是伴侣与上游聚集倾向结构域的相互作用是否增强了GFP荧光。在这里，我们证明，只有当识别聚集倾向蛋白并帮助其折叠的适当伴侣共表达时，GFP荧光才会显著增加。提出了GFP荧光与伴侣产物折叠之间的可能关联。这项研究可能为鉴定难折叠蛋白的特异性伴侣提供一种通用策略。
Green fluorescent protein (GFP) has been used to report protein folding by correlating solubility with fluorescence. In a GFP fusion protein, an upstream aggregation-prone domain can disrupt de novo folding of the GFP domain in Escherichia coli, resulting in a loss of fluorescence. Previously, we showed that prevention of misfolding of the upstream aggregation-prone domain by a coupled folding and binding interaction during protein synthesis restored both GFP fluorescence and solubility. Since molecular chaperones often fold nascent polypeptides through a bind-and-release interaction, the question remains whether the chaperone interaction with the upstream aggregation-prone domain enhances GFP fluorescence. Here, we demonstrate that a significant increase in GFP fluorescence occurred only when appropriate chaperones that recognized the aggregation-prone protein and helped its folding were co-expressed. A possible correlation between GFP fluorescence and the productive folding by chaperones is proposed. This study may provide a general strategy for identifying chaperones specific for difficult-to-fold proteins.