Purification and characterization of different proteasome species from mammalian cells.
| Autor: | Byun I; Department of Biochemistry and Molecular Biology, Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul 03080, Korea., Seo H; Department of Transdisciplinary Medicine, Seoul National University Hospital, Seoul 03080, Korea., Kim J; Department of Biochemistry and Molecular Biology, Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul 03080, Korea., Jeong D; Department of Biochemistry and Molecular Biology, Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul 03080, Korea., Han D; Department of Transdisciplinary Medicine, Seoul National University Hospital, Seoul 03080, Korea; Department of Medicine, Seoul National University College of Medicine, Seoul 03080, Korea. Electronic address: hdh03@snu.ac.kr., Lee MJ; Department of Biochemistry and Molecular Biology, Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul 03080, Korea. Electronic address: minjlee@snu.ac.kr. |
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| Jazyk: | angličtina |
| Zdroj: | STAR protocols [STAR Protoc] 2023 Dec 15; Vol. 4 (4), pp. 102748. Date of Electronic Publication: 2023 Nov 23. |
| DOI: | 10.1016/j.xpro.2023.102748 |
| Abstrakt: | Proteasomes are heterogeneous in forms and functions, but how the equilibrium among the 20S, 26S, and 30S proteasomes is achieved and altered is elusive. Here, we present a protocol for purifying and characterizing proteasome species. We describe steps for generating stable cell lines; affinity purifying the proteasome species; and characterizing them through native PAGE, activity assay, size-exclusion chromatography, and mass spectrometry. These standardized methods may contribute to biochemical studies of cellular proteasomes under both physiological and pathological conditions. For complete details on the use and execution of this protocol, please refer to Choi et al. (2023). 1 . Competing Interests: Declaration of interests The authors declare no competing interests. (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.) |
| Databáze: | MEDLINE |
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