Use of the LC3B-fusion technique for biochemical and structural studies of proteins involved in the N-degron pathway
DC Field | Value | Language |
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dc.contributor.author | Kim, Leehyeon | - |
dc.contributor.author | Kwon, Do Hoon | - |
dc.contributor.author | Heo, Jiwon | - |
dc.contributor.author | Park, Mi Rae | - |
dc.contributor.author | Song, Hyun Kyu | - |
dc.date.accessioned | 2021-08-31T09:35:44Z | - |
dc.date.available | 2021-08-31T09:35:44Z | - |
dc.date.created | 2021-06-18 | - |
dc.date.issued | 2020-02-28 | - |
dc.identifier.issn | 0021-9258 | - |
dc.identifier.uri | https://scholar.korea.ac.kr/handle/2021.sw.korea/57602 | - |
dc.description.abstract | The N-degron pathway, formerly the N-end rule pathway, is a protein degradation process that determines the half-life of proteins based on their N-terminal residues. In contrast to the well-established in vivo studies over decades, in vitro studies of this pathway, including biochemical characterization and high-resolution structures, are relatively limited. In this study, we have developed a unique fusion technique using microtubule-associated protein 1A/1B light chain 3B, a key marker protein of autophagy, to tag the N terminus of the proteins involved in the N-degron pathway, which enables high yield of homogeneous target proteins with variable N-terminal residues for diverse biochemical studies including enzymatic and binding assays and substrate identification. Intriguingly, crystallization showed a markedly enhanced probability, even for the N-degron complexes. To validate our results, we determined the structures of select proteins in the N-degron pathway and compared them with the Protein Data Bank?deposited proteins. Furthermore, several biochemical applications of this technique were introduced. Therefore, this technique can be used as a general tool for the in vitro study of the N-degron pathway. | - |
dc.language | English | - |
dc.language.iso | en | - |
dc.publisher | AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC | - |
dc.subject | END RULE PATHWAY | - |
dc.subject | SUMO FUSION TECHNOLOGY | - |
dc.subject | TERMINAL AMIDASE | - |
dc.subject | SUBSTRATE RECOGNITION | - |
dc.subject | UBIQUITIN LIGASE | - |
dc.subject | EXPRESSION | - |
dc.subject | COMPONENT | - |
dc.subject | SPECIFICITY | - |
dc.subject | PHOSPHORYLATION | - |
dc.subject | AUTOPHAGY | - |
dc.title | Use of the LC3B-fusion technique for biochemical and structural studies of proteins involved in the N-degron pathway | - |
dc.type | Article | - |
dc.contributor.affiliatedAuthor | Song, Hyun Kyu | - |
dc.identifier.doi | 10.1074/jbc.RA119.010912 | - |
dc.identifier.scopusid | 2-s2.0-85080865981 | - |
dc.identifier.wosid | 000519969100006 | - |
dc.identifier.bibliographicCitation | JOURNAL OF BIOLOGICAL CHEMISTRY, v.295, no.9, pp.2590 - 2600 | - |
dc.relation.isPartOf | JOURNAL OF BIOLOGICAL CHEMISTRY | - |
dc.citation.title | JOURNAL OF BIOLOGICAL CHEMISTRY | - |
dc.citation.volume | 295 | - |
dc.citation.number | 9 | - |
dc.citation.startPage | 2590 | - |
dc.citation.endPage | 2600 | - |
dc.type.rims | ART | - |
dc.type.docType | Article | - |
dc.description.journalClass | 1 | - |
dc.description.journalRegisteredClass | scie | - |
dc.description.journalRegisteredClass | scopus | - |
dc.relation.journalResearchArea | Biochemistry & Molecular Biology | - |
dc.relation.journalWebOfScienceCategory | Biochemistry & Molecular Biology | - |
dc.subject.keywordPlus | END RULE PATHWAY | - |
dc.subject.keywordPlus | SUMO FUSION TECHNOLOGY | - |
dc.subject.keywordPlus | TERMINAL AMIDASE | - |
dc.subject.keywordPlus | SUBSTRATE RECOGNITION | - |
dc.subject.keywordPlus | UBIQUITIN LIGASE | - |
dc.subject.keywordPlus | EXPRESSION | - |
dc.subject.keywordPlus | COMPONENT | - |
dc.subject.keywordPlus | SPECIFICITY | - |
dc.subject.keywordPlus | PHOSPHORYLATION | - |
dc.subject.keywordPlus | AUTOPHAGY | - |
dc.subject.keywordAuthor | protein crystallization | - |
dc.subject.keywordAuthor | ubiquitylation (ubiquitination) | - |
dc.subject.keywordAuthor | protein degradation | - |
dc.subject.keywordAuthor | autophagy | - |
dc.subject.keywordAuthor | p62 (sequestosome 1(SQSTM1)) | - |
dc.subject.keywordAuthor | LC3B | - |
dc.subject.keywordAuthor | N-end rule | - |
dc.subject.keywordAuthor | NTAQ1 | - |
dc.subject.keywordAuthor | PRT1 | - |
dc.subject.keywordAuthor | UBR box | - |
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