Simple visualization of submicroscopic protein clusters with a phase-separation-based fluorescent reporter
Copyright © 2024 Elsevier Inc. All rights reserved..
Protein clustering plays numerous roles in cell physiology and disease. However, protein oligomers can be difficult to detect because they are often too small to appear as puncta in conventional fluorescence microscopy. Here, we describe a fluorescent reporter strategy that detects protein clusters with high sensitivity called CluMPS (clusters magnified by phase separation). A CluMPS reporter detects and visually amplifies even small clusters of a binding partner, generating large, quantifiable fluorescence condensates. We use computational modeling and optogenetic clustering to demonstrate that CluMPS can detect small oligomers and behaves rationally according to key system parameters. CluMPS detected small aggregates of pathological proteins where the corresponding GFP fusions appeared diffuse. CluMPS also detected and tracked clusters of unmodified and tagged endogenous proteins, and orthogonal CluMPS probes could be multiplexed in cells. CluMPS provides a powerful yet straightforward approach to observe higher-order protein assembly in its native cellular context. A record of this paper's transparent peer review process is included in the supplemental information.
Medienart: |
E-Artikel |
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Erscheinungsjahr: |
2024 |
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Erschienen: |
2024 |
Enthalten in: |
Zur Gesamtaufnahme - volume:15 |
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Enthalten in: |
Cell systems - 15(2024), 2 vom: 21. Feb., Seite 166-179.e7 |
Sprache: |
Englisch |
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Beteiligte Personen: |
Mumford, Thomas R [VerfasserIn] |
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Links: |
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Themen: |
Clusters |
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Anmerkungen: |
Date Completed 26.02.2024 Date Revised 20.03.2024 published: Print-Electronic Citation Status MEDLINE |
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doi: |
10.1016/j.cels.2024.01.005 |
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funding: |
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Förderinstitution / Projekttitel: |
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PPN (Katalog-ID): |
NLM368254437 |
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520 | |a Protein clustering plays numerous roles in cell physiology and disease. However, protein oligomers can be difficult to detect because they are often too small to appear as puncta in conventional fluorescence microscopy. Here, we describe a fluorescent reporter strategy that detects protein clusters with high sensitivity called CluMPS (clusters magnified by phase separation). A CluMPS reporter detects and visually amplifies even small clusters of a binding partner, generating large, quantifiable fluorescence condensates. We use computational modeling and optogenetic clustering to demonstrate that CluMPS can detect small oligomers and behaves rationally according to key system parameters. CluMPS detected small aggregates of pathological proteins where the corresponding GFP fusions appeared diffuse. CluMPS also detected and tracked clusters of unmodified and tagged endogenous proteins, and orthogonal CluMPS probes could be multiplexed in cells. CluMPS provides a powerful yet straightforward approach to observe higher-order protein assembly in its native cellular context. A record of this paper's transparent peer review process is included in the supplemental information | ||
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700 | 1 | |a Rae, Diarmid |e verfasserin |4 aut | |
700 | 1 | |a Brackhahn, Emily |e verfasserin |4 aut | |
700 | 1 | |a Idris, Abbas |e verfasserin |4 aut | |
700 | 1 | |a Gonzalez-Martinez, David |e verfasserin |4 aut | |
700 | 1 | |a Pal, Ayush Aditya |e verfasserin |4 aut | |
700 | 1 | |a Chung, Michael C |e verfasserin |4 aut | |
700 | 1 | |a Guan, Juan |e verfasserin |4 aut | |
700 | 1 | |a Rhoades, Elizabeth |e verfasserin |4 aut | |
700 | 1 | |a Bugaj, Lukasz J |e verfasserin |4 aut | |
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