Highly chromophoric fluorescent-labeled methionyl-initiator tRNAs applicable in living cells

We present an optimized strategy to effectively generate highly fluorescent initiator-tRNA complexes in living cells. Our strategy allows the generation of Fluo-Met-tRNAiMet complexes. These complexes can have highly chromogenic N-terminal labeling. For generating such complexes, we use either purified fluorescent methionine (PFM) or non-purified fluorescently labeled methionine (NPFM). Furthermore, PFM promotes the active generation of endogenous tRNAi in cells, leading to highly efficient Fluo-Met-tRNAiMet complexes. Finally, PFM-tRNAiMet complexes also facilitate the visualization of native fluorescently labeled Tat binding to beads. This demonstrates the potential of our approach to advance precision protein engineering and biotechnology applications.PMID:38494424 | DOI:10.1002/biot.202300579

Source: Biotechnology Journal - March 17, 2024 Category: Biotechnology Authors: Jung Min Kim Jinho Jung Source Type: research