An easy tool to monitor the elemental steps of in vitro translation via gel electrophoresis of fluorescently labelled small peptides

Авторы: Marina Valeriya I., Bidzhieva Medina, Tereshchenkov Andrey G., Orekhov Dmitry, Sagitova Vladislava E., Sumbatyan Nataliya V., Tashlitsky Vadim N., Ferberg Artem S., Maviza Tinashe P., Kasatsky Pavel, Tolicheva Olga, Paleskava Alena, Polshakov Vladimir I., Osterman Ilya A., Dontsova Olga A., Konevega Andrey L., Sergiev Petr V.
Журнал: RNA
Год издания: 2024
Номер статьи: rna.079766.123
DOI: 10.1261/rna.079766.123
Аннотация: 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 to 7 amino acids, BODIPY-labelled peptides, can be monitored using urea-polyacrylamide gel electrophoresis. Very short BODIPY-labelled 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.
Добавил в систему: Марина Валерия Ивановна

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