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Highly chromophoric fluorescent-labeled methionyl-initiator tRNAs applicable in living cells.
- Source :
-
Biotechnology journal [Biotechnol J] 2024 Mar; Vol. 19 (3), pp. e2300579. Date of Electronic Publication: 2024 Mar 17. - Publication Year :
- 2024
-
Abstract
- Fluorescent initiator tRNAs (tRNAi) play a crucial role in studying protein synthesis, yet generating highly fluorescent tRNAi complexes remains challenging. We present an optimized strategy to effectively generate highly fluorescent initiator-tRNA complexes in living cells. Our strategy allows the generation of Fluo-Met-tRNAi <superscript>Met</superscript> 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-tRNAi <superscript>Met</superscript> complexes. Finally, PFM-tRNAi <superscript>Met</superscript> 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.<br /> (© 2024 The Authors. Biotechnology Journal published by Wiley‐VCH GmbH.)
Details
- Language :
- English
- ISSN :
- 1860-7314
- Volume :
- 19
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- Biotechnology journal
- Publication Type :
- Academic Journal
- Accession number :
- 38494424
- Full Text :
- https://doi.org/10.1002/biot.202300579