Mechanically transduced immunosorbent assay to measure protein-protein interactions

eLife

Brian D Strahl¹  Christopher J Petell¹  Michael Pappas²  Diego Sandoval²  Joseph S Harrison³  Kathyrn Randene³  Joshua P Steimel⁴ 

[1] Department of Biochemistry and Biophysics, The University of North Carolina School of Medicine, Chapel Hill, United States;UNC Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, United States;Department of Biological Engineering, University of the Pacific, Stockton, United States;Department of Chemistry, University of the Pacific, Stockton, United States;Department of Mechanical Engineering, University of the Pacific, Stockton, United States;

关键词: METRIS;    protein-protein interactions;    epigenetics;    ubiquitin;    UHRF1;    DIDO1;    None;   

DOI  :  10.7554/eLife.67525

来源: eLife Sciences Publications, Ltd

【 摘 要 】

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.

【 授权许可】

CC BY