Potentiation of the activity of Escherichia coli chaperone DnaJ by tailing hyper-acidic minipeptides
Potentiation of the activity of Escherichia coli chaperone DnaJ by tailing hyper-acidic minipeptides
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通过加尾超酸性小肽增强大肠杆菌分子伴侣 DnaJ 的活性
DOI：
10.1016/j.jbiotec.2021.09.012
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发表时间：
2021-09-30
影响因子：
4.1
通讯作者：
Zou, Zhurong
中科院分区：
文献类型：
--
作者：
Liu, Yanjuan;Zhang, Mengru;Zou, Zhurong
分子伴侣网络在细胞蛋白质稳态中起着重要作用。然而，一些核心组分通常与错误折叠的蛋白质共聚集以用于螯合和功能障碍，导致异常的细胞蛋白质稳定、聚集相关病症和过表达的重组蛋白质的溶解性差。其中，DnaJ或其直向同源物是DnaK-DnaJ-GrpE三元系统中的一种专性共分子伴侣，可能是由于其内在的聚集倾向。在此，我们通过使用超酸化蛋白融合策略来增强大肠杆菌DnaJ的活性。我们发现DnaJ确实只有中等的溶解度，可以通过融合超酸性小肽来显着改善。最重要的是，我们揭示了具有融合尾的高度酸化的DnaJ可以优于其天然形式（显著高达2.1倍），以提高靶蛋白的溶解度，同时适当地赋予它们升高的活性。这些结果表明，超酸化的DnaJ可以陪伴靶蛋白正确折叠成真正可溶的活性形式。此外，我们发现这些高度酸化的DnaJ变体可以超越其原型赋予E。大肠杆菌或酵母的耐热性增强，DnaJ本身可以被其过酸化的融合同源物溶解。最后，我们讨论了整体机制的DnaJ活性增强介导的超酸性拖尾融合。
The chaperone network plays an essential role in cellular protein homeostasis. However, some core components often coaggregate with misfolded proteins for sequestration and dysfunction, leading to abnormal cell proteostasis, aggregation-associated disorders, and poor solubility of overexpressed recombinant proteins. Among them, DnaJ or its ortholog, an obligate co-chaperone in the tripartite DnaK-DnaJ-GrpE system, is of more implications, probably due to its intrinsic propensity for aggregation. Herein, we potentiated the activity of Escherichia coli DnaJ by using hyper-acidified protein fusion strategy. We found DnaJ did possess only a moderate solubility that could be remarkably improved by fusing hyper-acidic minipeptides. Most importantly, we revealed the hyper-acidified DnaJ with a fusion tail could outperform its native form (significantly up to 2.1-fold) to enhance the solubility of target proteins and meanwhile appropriately impart them an elevated activity. These results suggest the hyper-acidified DnaJs can chaperone target proteins with correct folding into a truly soluble and active form. Moreover, we showed these hyper-acidified DnaJ variants could surpass its prototype to confer E. coli or yeast an enhanced heat tolerance, and DnaJ itself could be solubilized by its hyper-acidified fusion cognates. Finally, we discussed the overall mechanism for DnaJ activity potentiation mediated by hyper-acidic tailing fusion.