{"created":"2023-06-20T15:48:07.022979+00:00","id":2641,"links":{},"metadata":{"_buckets":{"deposit":"aaefa8d9-e4d2-4004-898b-109e18d3d06f"},"_deposit":{"created_by":4,"id":"2641","owners":[4],"pid":{"revision_id":0,"type":"depid","value":"2641"},"status":"published"},"_oai":{"id":"oai:hama-med.repo.nii.ac.jp:00002641","sets":["2:15:17"]},"author_link":["7772"],"item_5_alternative_title_1":{"attribute_name":"その他のタイトル","attribute_value_mlt":[{"subitem_alternative_title":"オプチニュリンオリゴマー：細胞内での主要な形態としての存在および酸化的刺激下でのユビキチン結合ドメインを介した架橋結合","subitem_alternative_title_language":"ja"}]},"item_5_biblio_info_5":{"attribute_name":"書誌情報","attribute_value_mlt":[{"bibliographicIssueDates":{"bibliographicIssueDate":"2013-09-20","bibliographicIssueDateType":"Issued"}}]},"item_5_date_granted_17":{"attribute_name":"学位授与年月日","attribute_value_mlt":[{"subitem_dategranted":"2013-09-20"}]},"item_5_degree_grantor_15":{"attribute_name":"学位授与機関","attribute_value_mlt":[{"subitem_degreegrantor":[{"subitem_degreegrantor_language":"ja","subitem_degreegrantor_name":"浜松医科大学"}],"subitem_degreegrantor_identifier":[{"subitem_degreegrantor_identifier_name":"13802","subitem_degreegrantor_identifier_scheme":"kakenhi"}]}]},"item_5_degree_name_12":{"attribute_name":"学位名","attribute_value_mlt":[{"subitem_degreename":"博士（医学）","subitem_degreename_language":"ja"}]},"item_5_description_13":{"attribute_name":"学位の区分","attribute_value_mlt":[{"subitem_description":"doctoral","subitem_description_language":"en","subitem_description_type":"Other"}]},"item_5_description_14":{"attribute_name":"学位の分野","attribute_value_mlt":[{"subitem_description":"医学系研究科","subitem_description_language":"ja","subitem_description_type":"Other"}]},"item_5_description_9":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"Optineurin gene, OPTN, is one of the causative genes of primary open-angle glaucoma. Although the oligomerization of optineurin in cultured cells was previously observed by gel filtration analysis and blue native gel electrophoresis (BNE), there is no detailed information about the feature of optineurin oligomer. The present study aims to analyze the oligomeric state of optineurin and its affecting factors, such as environmental stimuli or the mutations of optineurin. Here, we demonstrated that both endogenous and transfected optineurin exist as oligomer, rather than monomer in NIH3T3 cells using BNE or immunoprecipitation followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). We also applied in situ proximity ligation assay to visualize the self-interaction of optineurin in fixed HeLaS3 cells and reveal that the optineurin oligomer was localized diffusely in cytoplasm. Optineurin oligomer was usually detected as a single band at the size equal to optineurin monomer by SDS-PAGE, while there was an additional protein band with larger size observed when cells were treated with H2O2 before cell lysis followed by SDS-PAGE. The protein complex with larger size was proved to be optineurin oligomer by immunoprecipitation and therefore called covalent optineurin oligomer. Interestingly, in most common glaucoma-associated mutation of optineurin, E50K expressing cells, the covalent oligomer was formed even without H2O2 stimulation. Anti-oxidants inhibited the formation of E50K-induced covalent oligomer in different degrees. A series of truncated constructs of optineurin was used to reveal that covalent oligomer might be optineurin trimer and the ubiquitin binding domain is essential for this trimer formation. Our results indicates that optineurin trimer might be the basic unit of the oligomer. The oligomeric state can be affected by many factors, like demonstrated here that some covalent bond(s) is induced by H2O2 or transfection of E50K, which provides a novel insight into E50K pathogenesis. Furthermore, the regulation of oligomeric state becomes important worth further studies.","subitem_description_language":"en","subitem_description_type":"Abstract"}]},"item_5_dissertation_number_18":{"attribute_name":"学位授与番号","attribute_value_mlt":[{"subitem_dissertationnumber":"甲第662号"}]},"item_5_version_type_32":{"attribute_name":"著者版フラグ","attribute_value_mlt":[{"subitem_version_resource":"http://purl.org/coar/version/c_ab4af688f83e57aa","subitem_version_type":"AM"}]},"item_access_right":{"attribute_name":"アクセス権","attribute_value_mlt":[{"subitem_access_right":"open access","subitem_access_right_uri":"http://purl.org/coar/access_right/c_abf2"}]},"item_creator":{"attribute_name":"著者","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{"creatorName":"Gao, Jie","creatorNameLang":"en"}],"nameIdentifiers":[{"nameIdentifier":"7772","nameIdentifierScheme":"WEKO"}]}]},"item_files":{"attribute_name":"ファイル情報","attribute_type":"file","attribute_value_mlt":[{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2018-08-27"}],"displaytype":"detail","filename":"DT_662ronbun.pdf","filesize":[{"value":"1.6 MB"}],"format":"application/pdf","licensetype":"license_note","mimetype":"application/pdf","url":{"label":"論文本文","objectType":"fulltext","url":"https://hama-med.repo.nii.ac.jp/record/2641/files/DT_662ronbun.pdf"},"version_id":"0a8e3511-1255-4e30-a76a-6d5999933169"}]},"item_language":{"attribute_name":"言語","attribute_value_mlt":[{"subitem_language":"eng"}]},"item_resource_type":{"attribute_name":"資源タイプ","attribute_value_mlt":[{"resourcetype":"doctoral thesis","resourceuri":"http://purl.org/coar/resource_type/c_db06"}]},"item_title":"The optineurin oligomer: its existence as a major form in cells and cross-linking upon oxidative stimuli through ubiquitin binding domain","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"The optineurin oligomer: its existence as a major form in cells and cross-linking upon oxidative stimuli through ubiquitin binding domain","subitem_title_language":"en"}]},"item_type_id":"5","owner":"4","path":["17"],"pubdate":{"attribute_name":"PubDate","attribute_value":"2015-10-06"},"publish_date":"2015-10-06","publish_status":"0","recid":"2641","relation_version_is_last":true,"title":["The optineurin oligomer: its existence as a major form in cells and cross-linking upon oxidative stimuli through ubiquitin binding domain"],"weko_creator_id":"4","weko_shared_id":-1},"updated":"2023-10-23T06:11:33.108295+00:00"}