Preparation of stable recombinant Osm1 noncovalently bound with flavin adenosine dinucleotide cofactor for structural study
DC Field | Value | Language |
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dc.contributor.author | Kim, Sunghwan | - |
dc.contributor.author | Park, Hyun Ho | - |
dc.date.available | 2019-05-28T01:36:26Z | - |
dc.date.issued | 2019-03 | - |
dc.identifier.issn | 2053-230X | - |
dc.identifier.issn | 2053-230X | - |
dc.identifier.uri | https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/18192 | - |
dc.description.abstract | Osm1, a soluble fumarate reductase from Saccharomyces cerevisiae, is localized in both the mitochondria and the endoplasmic reticulum (ER). OSM1 genetically interacts with ERO1, which encodes an essential ER oxidoreductase for disulfide-bond formation under anaerobic conditions. However, the detailed enzymatic mechanisms involved in this interaction and the cellular roles of Osm1 are not fully understood. In this study, monomeric and stable recombinant Osm1 was successfully prepared for structural study. During purification, it was realized that the majority of recombinant Osm1 expressed in Escherichia coli lacked the flavin adenosine dinucleotide (FAD) cofactor. However, exogenously introduced FAD could be incorporated into recombinant Osm1, generating stable and homogenous holo Osm1. Moreover, after removing a flexible fragment by limited proteolysis, holo Osm1 formed isotropic crystals that retained catalytic activity. X-ray diffraction data were successfully collected from the Osm1 crystals to a resolution of 1.75 angstrom. | - |
dc.format.extent | 7 | - |
dc.language | 영어 | - |
dc.language.iso | ENG | - |
dc.publisher | John Wiley and Sons Ltd | - |
dc.title | Preparation of stable recombinant Osm1 noncovalently bound with flavin adenosine dinucleotide cofactor for structural study | - |
dc.type | Article | - |
dc.identifier.doi | 10.1107/S2053230X19000190 | - |
dc.identifier.bibliographicCitation | Acta Crystallographica Section F: Structural Biology Communications, v.75, pp 159 - 165 | - |
dc.description.isOpenAccess | N | - |
dc.identifier.wosid | 000460520900004 | - |
dc.identifier.scopusid | 2-s2.0-85062599018 | - |
dc.citation.endPage | 165 | - |
dc.citation.startPage | 159 | - |
dc.citation.title | Acta Crystallographica Section F: Structural Biology Communications | - |
dc.citation.volume | 75 | - |
dc.type.docType | Article | - |
dc.publisher.location | 영국 | - |
dc.subject.keywordAuthor | soluble fumarate reductase | - |
dc.subject.keywordAuthor | anaerobiosis | - |
dc.subject.keywordAuthor | Osm1 | - |
dc.subject.keywordAuthor | limited proteolysis | - |
dc.subject.keywordAuthor | stabilization | - |
dc.subject.keywordAuthor | crystallization | - |
dc.subject.keywordPlus | SOLUBLE FUMARATE REDUCTASE | - |
dc.subject.keywordPlus | SACCHAROMYCES-CEREVISIAE | - |
dc.subject.keywordPlus | ISOENZYMES | - |
dc.relation.journalResearchArea | Biochemistry & Molecular Biology | - |
dc.relation.journalResearchArea | Biophysics | - |
dc.relation.journalResearchArea | Crystallography | - |
dc.relation.journalWebOfScienceCategory | Biochemical Research Methods | - |
dc.relation.journalWebOfScienceCategory | Biochemistry & Molecular Biology | - |
dc.relation.journalWebOfScienceCategory | Biophysics | - |
dc.relation.journalWebOfScienceCategory | Crystallography | - |
dc.description.journalRegisteredClass | scie | - |
dc.description.journalRegisteredClass | scopus | - |
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