Mechanically transduced immunosorbent assay to measure protein-protein interactions.

DIDO1 METRIS UHRF1 biochemistry chemical biology epigenetics none physics of living systems protein-protein interactions ubiquitin

Journal

eLife
ISSN: 2050-084X
Titre abrégé: Elife
Pays: England
ID NLM: 101579614

Informations de publication

Date de publication:
28 09 2021
Historique:
received: 18 02 2021
accepted: 28 08 2021
entrez: 28 9 2021
pubmed: 29 9 2021
medline: 9 11 2021
Statut: epublish

Résumé

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.

Identifiants

pubmed: 34581668
doi: 10.7554/eLife.67525
pii: 67525
pmc: PMC8479797
doi:
pii:

Substances chimiques

Immunosorbents 0
Proteins 0

Types de publication

Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Subventions

Organisme : NIGMS NIH HHS
ID : R35 GM126900
Pays : United States

Informations de copyright

© 2021, Petell et al.

Déclaration de conflit d'intérêts

CP, KR, MP, DS, BS, JH No competing interests declared, JS is a cofounder of Tribosense Technologies

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Auteurs

Christopher J Petell (CJ)

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.

Kathyrn Randene (K)

Department of Chemistry, University of the Pacific, Stockton, United States.

Michael Pappas (M)

Department of Biological Engineering, University of the Pacific, Stockton, United States.

Diego Sandoval (D)

Department of Biological Engineering, University of the Pacific, Stockton, United States.

Brian D Strahl (BD)

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.

Joseph S Harrison (JS)

Department of Chemistry, University of the Pacific, Stockton, United States.

Joshua P Steimel (JP)

Department of Mechanical Engineering, University of the Pacific, Stockton, United States.

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Classifications MeSH