A High-Throughput Assay to Identify Specific Nascent Chain Inhibitors
Fischer, P. D.; Hiller, S. · biochemistry · 2026-09-06 · 原文
DOI：10.64898/2026.09.03.749102作者：2 位
"Undruggable" proteins without surface-accessible binding sites pose significant challenges to target-based drug discovery. Innovative approaches are needed to tackle these proteins. One promising strategy is targeting them in their nascent chain form at the ribosome, where they have a different conformation than in the folded form. A systematic approach to screen for such compounds has however been lacking. Here, we present a high-throughput assay to identify small molecules that specifically inhibit a protein of interest in its nascent-chain form. The assay employs a human in vitro transcription/translation system and monitors expression of the protein in real time via fluorescence detection. Specific inhibitors for the nascent chain of interest can then be identified by comparison with a counter screen. The assay was optimized to maximal sensitivity and reaction costs of ~$0.01 per well, enabling large-scale screens. We validated performance with the reference compound PF846 on the nascent chain of the protein PCSK9. Feasibility for high-throughput screening was demonstrated using a library of 1,760 compounds against the oncogenic KRAS variant A146T and the protein ApoC3, with m
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1. 人话版
"Undruggable" proteins without surface-accessible binding sites pose significant challenges to target-based drug discovery.
Innovative approaches are needed to tackle these proteins.
2. 领域脉络
本文类目：biochemistry，属于其所在研究脉络的最新进展。
3. 机制拆解
One promising strategy is targeting them in their nascent chain form at the ribosome, where they have a different conformation than in the folded form.
Here, we present a high-throughput assay to identify small molecules that specifically inhibit a protein of interest in its nascent-chain form.
4. 证据与数字
The assay was optimized to maximal sensitivity and reaction costs of ~$0.01 per well, enabling large-scale screens.
We validated performance with the reference compound PF846 on the nascent chain of the protein PCSK9.
Feasibility for high-throughput screening was demonstrated using a library of 1,760 compounds against the oncogenic KRAS variant A146T and the protein ApoC3, with m
5. 反例与边界
A systematic approach to screen for such compounds has however been lacking.
6. 跨领域连接与意外收获
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7. 可复用方法
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