生物
紧身衣
翻译(生物学)
凝胶电泳
聚丙烯酰胺凝胶电泳
肽
生物化学
氨基酸
寡肽
体外
转移RNA
电泳
色谱法
分子生物学
组合化学
荧光
信使核糖核酸
化学
基因
核糖核酸
酶
物理
量子力学
作者
V. I. Marina,Medina Bidzhieva,Andrey G. Tereshchenkov,Dmitry Orekhov,Vladislava E. Sagitova,N. V. Sumbatyan,Vadim N. Tashlitsky,Artem S. Ferberg,Tinashe P. Maviza,Pavel Kasatsky,Olga A Tolicheva,Alena Paleskava,Vladimir I. Polshakov,Ilya А. Osterman,Olga А. Dontsova,Andrey L. Konevega,Петр В. Сергиев
出处
期刊:RNA
日期:2023-12-26
卷期号:30 (3): 298-307
标识
DOI:10.1261/rna.079766.123
摘要
Several methods are available to visualize and assess the kinetics and efficiency of elemental steps of protein biosynthesis. However, each of these methods has its own limitations. Here, we present a novel, simple and convenient tool for monitoring stepwise in vitro translation initiated by BODIPY-Met-tRNA. Synthesis and release of very short, 1-7 amino acids, BODIPY-labeled peptides, can be monitored using urea-polyacrylamide gel electrophoresis. Very short BODIPY-labeled oligopeptides might be resolved this way, in contrast to widely used Tris-tricine gel electrophoresis, which is suitable to separate peptides larger than 1 kDa. The method described in this manuscript allows one to monitor the steps of translation initiation, peptide transfer, translocation, and termination as well as their inhibition at an unprecedented single amino acid resolution.
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