Mechanically transduced immunosorbent assay to measure protein-protein interactions
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 leve...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
eLife Sciences Publications Ltd
2021-09-01
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| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/67525 |
| _version_ | 1818018760702820352 |
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| author | Christopher J Petell Kathyrn Randene Michael Pappas Diego Sandoval Brian D Strahl Joseph S Harrison Joshua P Steimel |
| author_facet | Christopher J Petell Kathyrn Randene Michael Pappas Diego Sandoval Brian D Strahl Joseph S Harrison Joshua P Steimel |
| author_sort | Christopher J Petell |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-14T07:43:05Z |
| format | Article |
| id | doaj.art-3244e8957e2b4851885fa8b644198db3 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-14T07:43:05Z |
| publishDate | 2021-09-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-3244e8957e2b4851885fa8b644198db32022-12-22T02:05:25ZengeLife Sciences Publications LtdeLife2050-084X2021-09-011010.7554/eLife.67525Mechanically transduced immunosorbent assay to measure protein-protein interactionsChristopher J Petell0Kathyrn Randene1Michael Pappas2Diego Sandoval3Brian D Strahl4https://orcid.org/0000-0002-4947-6259Joseph S Harrison5https://orcid.org/0000-0002-2118-6524Joshua P Steimel6https://orcid.org/0000-0003-2437-8545Department 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 StatesDepartment of Chemistry, University of the Pacific, Stockton, United StatesDepartment of Biological Engineering, University of the Pacific, Stockton, United StatesDepartment of Biological Engineering, University of the Pacific, Stockton, United StatesDepartment 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 StatesDepartment of Chemistry, University of the Pacific, Stockton, United StatesDepartment of Mechanical Engineering, University of the Pacific, Stockton, United StatesMeasuring 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.https://elifesciences.org/articles/67525METRISprotein-protein interactionsepigeneticsubiquitinUHRF1DIDO1 |
| spellingShingle | Christopher J Petell Kathyrn Randene Michael Pappas Diego Sandoval Brian D Strahl Joseph S Harrison Joshua P Steimel Mechanically transduced immunosorbent assay to measure protein-protein interactions eLife METRIS protein-protein interactions epigenetics ubiquitin UHRF1 DIDO1 |
| title | Mechanically transduced immunosorbent assay to measure protein-protein interactions |
| title_full | Mechanically transduced immunosorbent assay to measure protein-protein interactions |
| title_fullStr | Mechanically transduced immunosorbent assay to measure protein-protein interactions |
| title_full_unstemmed | Mechanically transduced immunosorbent assay to measure protein-protein interactions |
| title_short | Mechanically transduced immunosorbent assay to measure protein-protein interactions |
| title_sort | mechanically transduced immunosorbent assay to measure protein protein interactions |
| topic | METRIS protein-protein interactions epigenetics ubiquitin UHRF1 DIDO1 |
| url | https://elifesciences.org/articles/67525 |
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