Slowing Bacterial Translation Speed Enhances Eukaryotic Protein Folding Efficiency
Slowing Bacterial Translation Speed Enhances Eukaryotic Protein Folding Efficiency
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DOI：
10.1016/j.jmb.2009.12.042
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发表时间：
2010-03-12
影响因子：
5.6
通讯作者：
Barral, Jose M.
中科院分区：
文献类型：
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作者：
Siller, Efrain;DeZwaan, Diane C.;Barral, Jose M.
细菌和真核生物之间的从头蛋白质折叠机制存在显着差异，这可以通过在细菌系统中重组生产时经常观察到的天然真核蛋白质的低产率来证明。细菌中的多肽合成速率比真核生物中的更快，但翻译速率的一般变化对蛋白质折叠效率的影响在很大程度上仍未被探索。通过使用具有可以调节翻译速度的突变核糖体的大肠杆菌细胞，我们在此表明降低多肽延伸率会导致真核来源的多种蛋白质的折叠增强。这些结果表明，在真核生物中，蛋白质折叠需要缓慢的翻译速率。相比之下，细菌中的折叠似乎与蛋白质合成无关，这解释了我们的发现，即翻译速度的普遍降低不会对内源细菌蛋白质组的折叠产生不利影响。利用这种策略可以生产出以前在细菌系统中无法获得的天然真核多结构域蛋白，并且可能成为生产易于聚集的重组蛋白的通用替代方案。 (C) 2009 Elsevier Ltd. 保留所有权利。
The mechanisms for de novo protein folding differ significantly between bacteria and eukaryotes, as evidenced by the often observed poor yields of native eukaryotic proteins upon recombinant production in bacterial systems. Polypeptide synthesis rates are faster in bacteria than in eukaryotes, but the effects of general variations in translation rates on protein folding efficiency have remained largely unexplored. By employing Escherichia coli cells with mutant ribosomes whose translation speed can be modulated, we show here that reducing polypeptide elongation rates leads to enhanced folding of diverse proteins of eukaryotic origin. These results suggest that in eukaryotes, protein folding necessitates slow translation rates. In contrast, folding in bacteria appears to be uncoupled from protein synthesis, explaining our findings that a generalized reduction in translation speed does not adversely impact the folding of the endogenous bacterial proteome. Utilization of this strategy has allowed the production of a native eukaryotic multidomain protein that has been previously unattainable in bacterial systems and may constitute a general alternative to the production of aggregation-prone recombinant proteins. (C) 2009 Elsevier Ltd. All rights reserved.