Cotranslational protein folding studies using Force Profile Analysis, a method where the SecM translational arrest peptide is used to detect folding‐induced forces acting on the nascent polypeptide, have so far been limited mainly to small domains of cytosolic proteins that fold in close proximity to the translating ribosome. In this study, we investigate the cotranslational folding of the periplasmic, disulfide bond‐containing Escherichia coli protein alkaline phosphatase (PhoA) in a wild‐type strain background and a strain background devoid of the periplasmic thiol: disulfide interchange protein DsbA. We find that folding‐induced forces can be transmitted via the nascent chain from the periplasm to the polypeptide transferase center in the ribosome, a distance of ~160 Å, and that PhoA appears to fold cotranslationally via at least two disulfide‐stabilized folding intermediates. Thus, Force Profile Analysis can be used to study cotranslational folding of proteins in an extra‐cytosolic compartment, like the periplasm.

中文翻译：

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大肠杆菌周质中碱性磷酸酶的共翻译折叠。


使用力分布分析（一种使用 SecM 翻译抑制肽来检测作用于新生多肽的折叠诱导力的方法）进行的共翻译蛋白质折叠研究，迄今为止主要限于折叠在紧邻蛋白质的胞浆蛋白的小结构域。翻译核糖体。在本研究中，我们研究了野生型菌株背景和不含周质硫醇：二硫键交换蛋白 DsbA 的菌株背景中含有二硫键的大肠杆菌蛋白碱性磷酸酶 (PhoA) 的共翻译折叠。我们发现折叠诱导力可以通过新生链从周质传递到核糖体中的多肽转移酶中心，距离约 160 Å，并且 PhoA 似乎通过至少两个二硫键稳定的折叠中间体进行共翻译折叠。因此，力分布分析可用于研究胞质外区室（如周质）中蛋白质的共翻译折叠。