Formation of active inclusion bodies in the periplasm of Escherichia coli
Formation of active inclusion bodies in the periplasm of Escherichia coli
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DOI：
10.1111/j.1365-2958.2006.05394.x
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发表时间：
2006-10-01
影响因子：
3.6
通讯作者：
Betton, Jean-Michel
中科院分区：
文献类型：
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作者：
Arie, Jean-Philippe;Miot, Marika;Betton, Jean-Michel
为了研究大肠杆菌外周质中折叠与聚集之间的关系，我们分析了输出蛋白与野生型麦芽糖结合蛋白（MalE）或易于聚集的变体MalE31融合的细胞命运。融合蛋白在周质中聚集的倾向是由上游蛋白固有的折叠特性决定的。当-内酰胺酶或碱性磷酸酶与MalE31的c端连接时，产生的融合蛋白以不溶性形式积累，但保持其催化活性。此外，这些蛋白聚集体诱导胞质外应激反应，类似于未融合的MalE31。然而，使用荧光底物，我们发现碱性磷酸酶活性存在于质周聚集体内。这些结果表明，质周包涵体的形成可能导致参与蛋白之间的分子间相互作用，而不会失去融合酶的功能。
To examine the relationship between folding and aggregation in the periplasm of Escherichia coli, we have analysed the cellular fates of exported proteins fused to either the wild-type maltose-binding protein (MalE) or the aggregation-prone variant MalE31. The propensity of fusion proteins to aggregate in the periplasm was determined by the intrinsic folding characteristics of the upstream protein. When beta-lactamase or alkaline phosphatase was linked to the C-terminus of MalE31, the resultant fusion proteins accumulated in an insoluble form, but retained their catalytic activity. In addition, these protein aggregates induced an extracytoplasmic stress response, similar to unfused MalE31. However, using a fluorescent substrate, we found that alkaline phosphatase activity was present inside periplasmic aggregates. These results suggest that periplasmic inclusion body formation may result in intermolecular interactions between participating proteins without loss of function of the fused enzymes.