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Updated: Feb 21, 2026
A Fluorescence-based Protocol for Preliminary Screening of Protein Synthesis Inhibitors from Natural Sources
Published on: January 27, 2026
High-Throughput Screening for Protein Synthesis Inhibitors Targeting Aminoacyl-tRNA Synthetases
Jiwon Kong1, Pengfei Fang2,3, Franck Madoux4,5
11 Medicinal Bioconvergence Research Center, College of Pharmacy, Seoul National University, Seoul, Korea.
Abstract:
Aminoacylation has been implicated in a wide variety of cancers. Aminoacyl-tRNA synthetases (ARSs) exist in large excess in tumor cells due to their increased demand for translation, whereas most other protein-synthesis apparatuses are quantitatively limited. Among other components that constitute the translation machinery-namely, tRNA, amino acid, ATP, and ARS-ARS is the only target that can be blocked by small molecules. No constitutively active ARSs have been reported, and mutations of ARS can cause inaccurate substrate recognition and malformation of the multi-ARS complex (MSC). Hence, interference of the activity is expected to be independent of genotype without developing resistance. Here, we report a high-throughput screening (HTS) system to find mammalian ARS inhibitors. The rabbit-reticulocyte lysate we used closely resembles both the individual and complexed structures of human ARSs, and it may predispose active compounds that are readily applicable for humankind. This assay was further validated because it identified familiar translational inhibitors from a pilot screen, such as emetine, proving its suitability for our purpose. The assay demonstrated excellent quality control (QC) parameters and reproducibility, and is proven ready for further HTS campaigns with large chemical libraries.
Insights
Researchers developed a high-throughput screening system to identify inhibitors of aminoacyl-tRNA synthetases (ARSs), crucial targets in cancer therapy. This validated assay is ready for large-scale screening to find novel anti-cancer drugs.
Area of Science:
- Biochemistry
- Molecular Biology
- Oncology
Background:
- Aminoacylation, catalyzed by aminoacyl-tRNA synthetases (ARSs), is vital for protein synthesis.
- ARSs are overexpressed in cancer cells, making them attractive therapeutic targets.
- Unlike other translation machinery components, ARSs can be inhibited by small molecules.
Purpose of the Study:
- To develop and validate a high-throughput screening (HTS) system for identifying mammalian ARS inhibitors.
- To establish a reliable assay for discovering novel anti-cancer compounds targeting ARSs.
Main Methods:
- A high-throughput screening (HTS) system was established using rabbit-reticulocyte lysate.
- The system was validated by identifying known translational inhibitors like emetine.
- Quality control and reproducibility parameters were assessed.
Main Results:
- The developed HTS system effectively mimics human ARS structures and function.
- The assay demonstrated excellent quality control and reproducibility.
- The system is suitable for large-scale screening of chemical libraries.
Conclusions:
- A robust HTS system for discovering mammalian ARS inhibitors has been successfully developed.
- This assay provides a valuable tool for identifying novel anti-cancer therapeutics targeting ARSs.
- The system's validation and readiness for HTS campaigns offer a promising avenue for drug discovery.
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