Proteomic analysis of nascent polypeptide chains that potentially induce translational pausing during elongation
Proteomic analysis of nascent polypeptide chains that potentially induce translational pausing during elongation
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DOI：
10.1093/bbb/zbac097
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发表时间：
2022-06-24
影响因子：
1.6
通讯作者：
Hayano，Toshiya
中科院分区：
文献类型：
--
作者：
Shimohata，Nobuyuki;Harada，Yudai;Hayano，Toshiya
目前，广泛研究了配备有“核糖体阻滞肽”（RAP）的蛋白质，所述蛋白质通过在延伸期间暂停其自身的翻译来调节下游基因的表达及其自身的活性。然而，RAP的研究主要是在原核细胞中进行的;对真核细胞，特别是哺乳动物细胞的研究有限。在本研究中，我们全面研究了在哺乳动物细胞中捕获的新生多肽，以获得对RAP的新见解。十六烷基三甲基溴化铵用于获得新生多肽链，其在翻译延伸期间被抑制。蛋白质组学分析后，通过根据C-末端的氨基酸残基进行鉴别的额外筛选揭示了几种新的RAP候选物。我们的方法可以应用于哺乳动物细胞中的全面RAP研究。
Currently, proteins equipped with “ribosomal arrest peptides” (RAPs) that regulate the expression of downstream genes and their own activity by pausing their own translation during elongation are extensively studied. However, studies focusing on RAP have been conducted primarily in prokaryotic cells; studies on eukaryotic cells, especially mammalian cells, are limited. In the present study, we comprehensively examined translationally arrested nascent polypeptides to gain novel insights into RAPs in mammalian cells. Cetyltrimethylammonium bromide was used to obtain nascent polypeptide chains that were translationally arrested during translation elongation. After proteomic analysis, additional screening by discriminating according to amino acid residues at the C-terminal end revealed several novel RAP candidates. Our method can be applied for comprehensive RAP studies in mammalian cells.