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Evaluation of Immunoproteasome-Specific Proteolytic Activity Using Fluorogenic Peptide Substrates
DC Field | Value | Language |
---|---|---|
dc.contributor.author | Kim, Sumin | - |
dc.contributor.author | Park, Seo Hyeong | - |
dc.contributor.author | Choi, Won Hoon | - |
dc.contributor.author | Lee, Min Jae | - |
dc.date.accessioned | 2022-10-05T04:11:07Z | - |
dc.date.available | 2022-10-05T04:11:07Z | - |
dc.date.created | 2022-07-21 | - |
dc.date.issued | 2022-06 | - |
dc.identifier.citation | Immune Network, Vol.22 No.3, p. e28 | - |
dc.identifier.issn | 1598-2629 | - |
dc.identifier.uri | https://hdl.handle.net/10371/185320 | - |
dc.description.abstract | The 26S proteasome irreversibly hydrolyzes polyubiquitylated substrates to maintain protein homeostasis; it also regulates immune responses by generating antigenic peptides. An alternative form of the 26S proteasome is the immunoproteasome, which contains substituted catalytic subunits (beta 1i/PSMB9, beta 2i/PSMB10, and beta 5i/PSMB8) instead of constitutively expressed counterparts (beta 1/PSMB6, beta 2/PSMB7, and beta 5/PSMB5). The immunoproteasome expands the peptide repertoire presented on MHC class I molecules. However, how its activity changes in this context is largely elusive, possibly due to the lack of a standardized methodology to evaluate its specific activity. Here, we describe an assay protocol that measures the immunoproteasome activity of whole-cell lysates using commercially available fluorogenic peptide substrates. Our results showed that the most accurate assessment of immunoproteasome activity could be achieved by combining beta 5i-targeting substrate Ac-ANW-AMC and immunoproteasome inhibitor ONX-0914. This simple and reliable protocol may contribute to future studies of immunoproteasomes and their pathophysiological roles during viral infection, inflammation, and tumorigenesis. | - |
dc.language | 영어 | - |
dc.publisher | 대한면역학회 | - |
dc.title | Evaluation of Immunoproteasome-Specific Proteolytic Activity Using Fluorogenic Peptide Substrates | - |
dc.type | Article | - |
dc.identifier.doi | 10.4110/in.2022.22.e28 | - |
dc.citation.journaltitle | Immune Network | - |
dc.identifier.wosid | 000821383900005 | - |
dc.identifier.scopusid | 2-s2.0-85133649557 | - |
dc.citation.number | 3 | - |
dc.citation.startpage | e28 | - |
dc.citation.volume | 22 | - |
dc.identifier.kciid | ART002858944 | - |
dc.description.isOpenAccess | N | - |
dc.contributor.affiliatedAuthor | Lee, Min Jae | - |
dc.type.docType | Article | - |
dc.description.journalClass | 1 | - |
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