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A biochemical reconstitution approach to identify autophagy receptors for aggrephagy in mammalian cells
Aggrephagy is a major way to clear protein aggregates. Here, we describe a pipeline of experiments to find autophagy receptors for aggrephagy. Steps include an in vitro reconstitution to recapitulate autophagosome recognizing aggregates and receptor identification steps based on flow cytometry and m...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9471455/ https://www.ncbi.nlm.nih.gov/pubmed/36097383 http://dx.doi.org/10.1016/j.xpro.2022.101662 |
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author | Ma, Xinyu Zhang, Wenhao Deng, Haiteng Zhang, Min Ge, Liang |
author_facet | Ma, Xinyu Zhang, Wenhao Deng, Haiteng Zhang, Min Ge, Liang |
author_sort | Ma, Xinyu |
collection | PubMed |
description | Aggrephagy is a major way to clear protein aggregates. Here, we describe a pipeline of experiments to find autophagy receptors for aggrephagy. Steps include an in vitro reconstitution to recapitulate autophagosome recognizing aggregates and receptor identification steps based on flow cytometry and mass spectrometry. We also describe functional validation steps based on immunofluorescence and immunoblot. The protocol provides a practical way to identify aggrephagy receptors. For complete details on the use and execution of this protocol, please refer to Ma et al. (2022). |
format | Online Article Text |
id | pubmed-9471455 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-94714552022-09-15 A biochemical reconstitution approach to identify autophagy receptors for aggrephagy in mammalian cells Ma, Xinyu Zhang, Wenhao Deng, Haiteng Zhang, Min Ge, Liang STAR Protoc Protocol Aggrephagy is a major way to clear protein aggregates. Here, we describe a pipeline of experiments to find autophagy receptors for aggrephagy. Steps include an in vitro reconstitution to recapitulate autophagosome recognizing aggregates and receptor identification steps based on flow cytometry and mass spectrometry. We also describe functional validation steps based on immunofluorescence and immunoblot. The protocol provides a practical way to identify aggrephagy receptors. For complete details on the use and execution of this protocol, please refer to Ma et al. (2022). Elsevier 2022-09-08 /pmc/articles/PMC9471455/ /pubmed/36097383 http://dx.doi.org/10.1016/j.xpro.2022.101662 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Protocol Ma, Xinyu Zhang, Wenhao Deng, Haiteng Zhang, Min Ge, Liang A biochemical reconstitution approach to identify autophagy receptors for aggrephagy in mammalian cells |
title | A biochemical reconstitution approach to identify autophagy receptors for aggrephagy in mammalian cells |
title_full | A biochemical reconstitution approach to identify autophagy receptors for aggrephagy in mammalian cells |
title_fullStr | A biochemical reconstitution approach to identify autophagy receptors for aggrephagy in mammalian cells |
title_full_unstemmed | A biochemical reconstitution approach to identify autophagy receptors for aggrephagy in mammalian cells |
title_short | A biochemical reconstitution approach to identify autophagy receptors for aggrephagy in mammalian cells |
title_sort | biochemical reconstitution approach to identify autophagy receptors for aggrephagy in mammalian cells |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9471455/ https://www.ncbi.nlm.nih.gov/pubmed/36097383 http://dx.doi.org/10.1016/j.xpro.2022.101662 |
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