Mechanically transduced immunosorbent assay to measure protein-protein interactions.
Mechanically transduced immunosorbent assay to measure protein-protein interactions.
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DOI：
10.7554/elife.67525
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发表时间：
2021-09-28
期刊：
影响因子：
7.7
通讯作者：
Steimel JP
中科院分区：
文献类型：
--
作者：
Petell CJ;Randene K;Pappas M;Sandoval D;Strahl BD;Harrison JS;Steimel JP
测定蛋白质-蛋白质相互作用（PPI）的亲和力是生物化学的基础。然而，传统的方法依赖于质量作用定律，由于试剂的缺乏和可测量亲和力范围的限制，无法测量许多ppi。在这里，我们提出了一种新的技术，利用摩擦的基本概念来产生与结合势相关的机械信号。机械转导免疫吸附（METRIS）试验利用滚动磁探针来测量PPI相互作用的亲和力。METRIS测量蛋白包被颗粒在蛋白功能化底物上的平移位移。平移位移与PPI引起的有效摩擦成比例，因此在发生结合事件时产生机械信号。METRIS检测使用少至20 mol的试剂来测量大范围的亲和力，同时表现出高分辨率和灵敏度。我们使用METRIS来测量以前使用传统方法无法获得的几种ppi，为表观遗传识别提供了新的见解。
Measuring protein-protein interaction (PPI) affinities is fundamental to biochemistry. Yet, conventional methods rely upon the law of mass action and cannot measure many PPIs due to a scarcity of reagents and limitations in the measurable affinity ranges. Here, we present a novel technique that leverages the fundamental concept of friction to produce a mechanical signal that correlates to binding potential. The mechanically transduced immunosorbent (METRIS) assay utilizes rolling magnetic probes to measure PPI interaction affinities. METRIS measures the translational displacement of protein-coated particles on a protein-functionalized substrate. The translational displacement scales with the effective friction induced by a PPI, thus producing a mechanical signal when a binding event occurs. The METRIS assay uses as little as 20 pmols of reagents to measure a wide range of affinities while exhibiting a high resolution and sensitivity. We use METRIS to measure several PPIs that were previously inaccessible using traditional methods, providing new insights into epigenetic recognition.