核糖体
释放系数
肽
蛋白质生物合成
大肠杆菌
核糖体蛋白
翻译(生物学)
核糖体RNA
生物
平移移码
化学
生物化学
信使核糖核酸
细胞生物学
核糖核酸
基因
作者
Teruyo Ojima‐Kato,Yuma Nishikawa,Yuki Furukawa,Takaaki Kojima,Hideo Nakano
标识
DOI:10.1016/j.jbc.2023.104676
摘要
The insertion of the DNA sequence encoding SKIK peptide adjacent to the M start codon of a difficult-to-express protein enhances protein production in Escherichia coli. In this report, we reveal that the increased production of the SKIK-tagged protein is not due to codon usage of the SKIK sequence. Furthermore, we found that insertion of SKIK or MSKIK just before the SecM arrest peptide (FSTPVWISQAQGIRAGP), which causes ribosomal stalling on mRNA, greatly increased the production of the protein containing the SecM arrest peptide in the E. coli-reconstituted cell-free protein synthesis system (PURE system). A similar translation enhancement phenomenon by MSKIK was observed for the CmlA leader peptide, a ribosome arrest peptide, whose arrest is induced by chloramphenicol. These results strongly suggest that the nascent MSKIK peptide prevents or releases ribosomal stalling immediately following its generation during the translation process, resulting in an increase of protein production.
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