{"created":"2023-07-27T06:55:03.724075+00:00","id":50719,"links":{},"metadata":{"_buckets":{"deposit":"b75394d0-b8af-439c-93a9-37efef8da3e6"},"_deposit":{"created_by":18,"id":"50719","owners":[18],"pid":{"revision_id":0,"type":"depid","value":"50719"},"status":"published"},"_oai":{"id":"oai:kanazawa-u.repo.nii.ac.jp:00050719","sets":["2812:2813:2826"]},"author_link":["24839","91837"],"item_9_biblio_info_8":{"attribute_name":"書誌情報","attribute_value_mlt":[{"bibliographicIssueDates":{"bibliographicIssueDate":"2008-05","bibliographicIssueDateType":"Issued"},"bibliographicPageStart":"9p.","bibliographicVolumeNumber":"2006-2007","bibliographic_titles":[{"bibliographic_title":"平成19(2007)年度 科学研究費補助金 基盤研究(C) 研究成果報告書"},{"bibliographic_title":"2007 Fiscal Year Final Research Report Summary","bibliographic_titleLang":"en"}]}]},"item_9_creator_33":{"attribute_name":"著者別表示","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{}],"nameIdentifiers":[{},{}]}]},"item_9_description_21":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"細胞表面受容体RAGEの細胞内シグナル生成に関わる分子を同定することを目的とし、研究実施計画に基づいて研究を遂行した結果、以下の成果を得た。\n1.RAGE分子のアスパラギン酸結合型糖鎖修飾部位に変異を導入し、糖鎖を持たないRAGE分子を細胞に発現させることにより、糖鎖のリガンド結合およびシグナル生成への関与を検討した。その結果、糖鎖の付加によって一部のリガンドとの親和性が低下し、細胞内シグナル生成も減弱していることが明らかとなった。\n2.RAGE分子の細胞外ドメインに対するモノクローナル抗体を蛍光色素で標識し、生細胞表面RAGEを蛍光標識することによりRAGE分子の挙動を観察した。その結果、RAGEはpinocytosisの関わる機構により恒常的に細胞内へ取り込まれていることが明らかとなった。この細胞内取り込みはリガンド刺激による影響を受けなかった。\n3.上記モノクローナル抗体をFRETのドナーとアクセプターの対をなす2種の蛍光色素で標識し、生細胞表面RAGE分子同士の2分子FRETを観察することにより、高分子リガンド依存性のRAGEのオリゴマー化の検出に成功した。\n4.RAGE分子のオリゴマー化を引き起こさないと予想される低分子量ヘパリンなどの低分子RAGEリガンドは、RAGEを介する細胞内シグナルを惹起せず、むしろ高分子リガンドによるシグナル生成を阻害する。\nこれらの新知見から、高分子リガンドによるRAGEのオリゴマー化が細胞内シグナル生成の最初のステップであることが強く示唆され、臨床的応用が期待されるRAGE拮抗薬開発の基本原理が明らかになった。","subitem_description_type":"Abstract"},{"subitem_description":"RAGE (receptor for advanced glycation endproducts) is a cell surface receptor that bind several ligands, such as AGE (advanced glycation endproducts), HMGB-1 (high mobility group box-I protein), amyloid beta. The intracellular signals evoked by RAGE-ligand interaction were thought to play pivotal roles in development of various diseases. However, the mechanism of generation of intracellular signals by RAGE was poorly understood.\nThis study shed lights on several new aspects of RAGE signaling mechanism as follows.\nI. Recombinant wild-type, de-N-glycosylation and G82S RAGE proteins were produced in COS-7 cells, purified and assayed for ligand-binding abilities. De-N-glycosylation at N81 and G82S mutation decreased Kd for glycolaldehyde-derived AGE to three orders of magnitude lower levels compared with wild-type. AGE-induced upregulation of VEGF mRNA was significantly augmented in endothelial cell-derived ECV304 cells expressing de-N-glycosylated and G82S RAGE when compared with wild-type expresser.\n2. RAGE molecules of living cell surface were labeled by fluorescent dye-conjugated monoclonal antibody that binds to extracellular domain of RAGE (mAbl3G4), and the dynamic state of RAGE was analyzed by confocal fluorescence laser microscopy. The results indicated that RAGE molecules were constitutively internalized, and pinocytotic mechanism was, at least partly, involved in RAGE internalization. The rate of RAGE internalization was not altered by addition of RAGE ligand.\n3. RAGE molecules of living cell surface were labeled by mixture of mAbl3G4 conjugated with Alexa 488 and mAbl3G4 with Alexa 594, which are a pair of donor and acceptor for FRET (fluorescence resonance energy transfer), to detect oligomerization of RAGE molecule by FRET. The results indicated that the RAGE molecule oligomerized upon binding its ligand.\n4. Low molecular weight RAGE ligands such as low molecular weight heparin, which are speculated not to induce the RAGE oligomerization, did not evoke RAGE-mediated intracellular signal, and inhibited the signaling by high molecular weight ligands.\nThese new findings strongly suggested that the oligomerization is the first step of the signal generation, and revealed the principle of the development of RAGE antagonists that should be promising candidates of the preventive medicine for RAGE-related diseases.","subitem_description_type":"Abstract"}]},"item_9_description_22":{"attribute_name":"内容記述","attribute_value_mlt":[{"subitem_description":"研究課題/領域番号:18590260, 研究期間(年度):2006-2007","subitem_description_type":"Other"},{"subitem_description":"出典：「マルチリガンド受容体RAGEによる細胞内シグナル生成機構の解明」研究成果報告書　課題番号18590260\n（KAKEN：科学研究費助成事業データベース（国立情報学研究所））\n（https://kaken.nii.ac.jp/report/KAKENHI-PROJECT-18590260/185902602007kenkyu_seika_hokoku_gaiyo/)を加工して作成","subitem_description_type":"Other"}]},"item_9_description_5":{"attribute_name":"提供者所属","attribute_value_mlt":[{"subitem_description":"金沢大学医学系研究科","subitem_description_type":"Other"}]},"item_9_identifier_registration":{"attribute_name":"ID登録","attribute_value_mlt":[{"subitem_identifier_reg_text":"10.24517/00057027","subitem_identifier_reg_type":"JaLC"}]},"item_9_relation_28":{"attribute_name":"関連URI","attribute_value_mlt":[{"subitem_relation_name":[{"subitem_relation_name_text":"https://kaken.nii.ac.jp/search/?qm=40303268"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/search/?qm=40303268","subitem_relation_type_select":"URI"}},{"subitem_relation_name":[{"subitem_relation_name_text":"https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18590260/"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18590260/","subitem_relation_type_select":"URI"}},{"subitem_relation_name":[{"subitem_relation_name_text":"https://kaken.nii.ac.jp/report/KAKENHI-PROJECT-18590260/185902602007kenkyu_seika_hokoku_gaiyo/"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/report/KAKENHI-PROJECT-18590260/185902602007kenkyu_seika_hokoku_gaiyo/","subitem_relation_type_select":"URI"}}]},"item_9_version_type_25":{"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_files":{"attribute_name":"ファイル情報","attribute_type":"file","attribute_value_mlt":[{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2021-05-14"}],"displaytype":"detail","filename":"ME-PR-WATANABE-T-kaken 2008-9p.pdf","filesize":[{"value":"394.7 kB"}],"format":"application/pdf","licensetype":"license_11","mimetype":"application/pdf","url":{"label":"ME-PR-WATANABE-T-kaken 2008-9p.pdf","url":"https://kanazawa-u.repo.nii.ac.jp/record/50719/files/ME-PR-WATANABE-T-kaken 2008-9p.pdf"},"version_id":"e7251e04-dd1b-420e-9b4d-b7faa33f0073"}]},"item_language":{"attribute_name":"言語","attribute_value_mlt":[{"subitem_language":"jpn"}]},"item_resource_type":{"attribute_name":"資源タイプ","attribute_value_mlt":[{"resourcetype":"research report","resourceuri":"http://purl.org/coar/resource_type/c_18ws"}]},"item_title":"マルチリガンド受容体RAGEによる細胞内シグナル生成機構の解明","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"マルチリガンド受容体RAGEによる細胞内シグナル生成機構の解明"},{"subitem_title":"Study on signal generation mechanism of a multi-ligand receptor,RAGE","subitem_title_language":"en"}]},"item_type_id":"9","owner":"18","path":["2826"],"pubdate":{"attribute_name":"公開日","attribute_value":"2021-05-14"},"publish_date":"2021-05-14","publish_status":"0","recid":"50719","relation_version_is_last":true,"title":["マルチリガンド受容体RAGEによる細胞内シグナル生成機構の解明"],"weko_creator_id":"18","weko_shared_id":-1},"updated":"2023-07-27T14:22:19.516966+00:00"}