4.8 Article

Mechanically transduced immunosorbent assay to measure protein-protein interactions

Journal

ELIFE
Volume 10, Issue -, Pages -

Publisher

eLIFE SCIENCES PUBL LTD
DOI: 10.7554/eLife.67525

Keywords

METRIS; protein-protein interactions; epigenetics; ubiquitin; UHRF1; DIDO1; None

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Funding

  1. University of the Pacific
  2. National Institute of General Medical Sciences [GM126900]
  3. American Cancer Society [PF-19-027- 459 01-DMC]

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The METRIS assay leverages the concept of friction to measure protein-protein interaction affinities, offering high resolution and sensitivity. It allows for the measurement of a wide range of affinities with a small amount of reagents, providing new insights into epigenetic recognition.
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.

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