Split-BioID a conditional proteomics approach to monitor the composition of spatiotemporally defined protein complexes
Understanding the function of the thousands of cellular proteins is a central question in molecular cell biology. As proteins are typically part of multiple dynamic and often overlapping macromolecular complexes exerting distinct functions, the identification of protein-protein interactions (PPI) and their assignment to specific complexes is a crucial but challenging task. We present a protein fragments complementation assay integrated with the proximity-dependent biotinylation technique BioID. Activated on the interaction of two proteins, split-BioID is a conditional proteomics approach that allows in a single and simple assay to both experimentally validate binary PPI and to unbiasedly identify additional interacting factors. Applying our method to the miRNA-mediated silencing pathway, we can probe the proteomes of two distinct functional complexes containing the Ago2 protein and uncover the protein GIGYF2 as a regulator of miRNA-mediated translation repression. Hence, we provide a novel tool to study dynamic spatiotemporally defined protein complexes in their native cellular environment.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2017 |
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| Erschienen: |
2017 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:8 |
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| Enthalten in: |
Nature communications - 8(2017) vom: 06. Juni, Seite 15690 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Schopp, Isabel Myriam [VerfasserIn] |
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| Links: |
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| Themen: |
Carrier Proteins |
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| Anmerkungen: |
Date Completed 19.11.2018 Date Revised 19.11.2018 published: Electronic Citation Status MEDLINE |
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| doi: |
10.1038/ncomms15690 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM272657816 |
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| 520 | |a Understanding the function of the thousands of cellular proteins is a central question in molecular cell biology. As proteins are typically part of multiple dynamic and often overlapping macromolecular complexes exerting distinct functions, the identification of protein-protein interactions (PPI) and their assignment to specific complexes is a crucial but challenging task. We present a protein fragments complementation assay integrated with the proximity-dependent biotinylation technique BioID. Activated on the interaction of two proteins, split-BioID is a conditional proteomics approach that allows in a single and simple assay to both experimentally validate binary PPI and to unbiasedly identify additional interacting factors. Applying our method to the miRNA-mediated silencing pathway, we can probe the proteomes of two distinct functional complexes containing the Ago2 protein and uncover the protein GIGYF2 as a regulator of miRNA-mediated translation repression. Hence, we provide a novel tool to study dynamic spatiotemporally defined protein complexes in their native cellular environment | ||
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| 700 | 1 | |a Amaya Ramirez, Cinthia Claudia |e verfasserin |4 aut | |
| 700 | 1 | |a Debeljak, Jerneja |e verfasserin |4 aut | |
| 700 | 1 | |a Kreibich, Elisa |e verfasserin |4 aut | |
| 700 | 1 | |a Skribbe, Merle |e verfasserin |4 aut | |
| 700 | 1 | |a Wild, Klemens |e verfasserin |4 aut | |
| 700 | 1 | |a Béthune, Julien |e verfasserin |4 aut | |
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