{"created":"2023-07-27T06:57:34.062868+00:00","id":54809,"links":{},"metadata":{"_buckets":{"deposit":"0d3652d5-9cca-4753-8c3f-310b33528263"},"_deposit":{"created_by":18,"id":"54809","owners":[18],"pid":{"revision_id":0,"type":"depid","value":"54809"},"status":"published"},"_oai":{"id":"oai:kanazawa-u.repo.nii.ac.jp:00054809","sets":["2812:2813:2828"]},"author_link":["62509"],"item_9_biblio_info_8":{"attribute_name":"書誌情報","attribute_value_mlt":[{"bibliographicIssueDates":{"bibliographicIssueDate":"2016-04-21","bibliographicIssueDateType":"Issued"},"bibliographicPageStart":"2p.","bibliographicVolumeNumber":"2003 – 2005","bibliographic_titles":[{"bibliographic_title":"平成17(2005)年度 科学研究費補助金 若手研究(B) 研究概要"},{"bibliographic_title":"2005 Research Project Summary","bibliographic_titleLang":"en"}]}]},"item_9_creator_33":{"attribute_name":"著者別表示","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{"creatorName":"Okishio, Nobuyuki"}],"nameIdentifiers":[{"nameIdentifier":"62509","nameIdentifierScheme":"WEKO"},{"nameIdentifier":"20262561","nameIdentifierScheme":"e-Rad","nameIdentifierURI":"https://kaken.nii.ac.jp/ja/search/?qm=20262561"},{"nameIdentifier":"20262561","nameIdentifierScheme":"研究者番号","nameIdentifierURI":"https://nrid.nii.ac.jp/nrid/1000020262561"}]}]},"item_9_description_21":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"アクチン重合を制御し、細胞骨格構築に関与するN-WASP [neural-WASP (Wiskott-Aldrich syndrome protein)]およびWAVE(WASP family verprolin-homologous protein)の活性制御メカニズムを明らかにするために、それらの調節因子WISH(WASP interacting SH3 protein)、Endophilin、IRSp53(insulin receptor substrate p53)のSH3ドメインに注目し、これらSH3ドメインの大腸菌を用いた大量発現・精製系の構築を試みた。いずれのSH3ドメインも、GSTタグを付加する発現プラスミドの構築には問題がなかった。しかしながら、WISH SH3では、発現タンパク質の大部分が不溶性であり、Endophilin SH3では、Cys残基を含むためか、発現タンパク質の一部が、非還元状態で二量体化しており、いずれの場合も、以後の実験に用いるには適さなかった。またEndophilin SH3では、Cys残基をSer残基に置換した変異体を作成したが、変異によって可溶性が低下するため、大量発現・精製には不向きであった。一方、IRSp53 SH3では、Cys残基を含まず、充分量の可溶性タンパクを得ることができた。ファージディスプレイライブラリー法を用いて、IRSp53 SH3が認識する配列を調べたところ、NRPVAPPLPFWLの配列を得た。CDを用いて、IRSp53 SH3と合成ペプチドAc-NRPV APPLPFWL-NH_2の親和性(Kd値)を調べたところ、約20-40μMであった。NMRを用いて、IRSp53 SH3の^<15>N-^1H HSQCスペクトルを測定したところ、残基間でシグナル強度の不均一が見られたため、残念ながら、構造決定を行うことができなかった。","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:15790147, 研究期間(年度):2003 – 2005","subitem_description_type":"Other"},{"subitem_description":"出典：「立体構造解析に基づくWASP・WAVEファミリータンパク質の機能制御」研究成果報告書　課題番号15790147\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/ja/grant/KAKENHI-PROJECT-15790147/)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":"金沢大学医薬保健研究域医学系","subitem_description_type":"Other"}]},"item_9_identifier_registration":{"attribute_name":"ID登録","attribute_value_mlt":[{"subitem_identifier_reg_text":"10.24517/00061085","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/ja/search/?kw=20262561"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/ja/search/?kw=20262561","subitem_relation_type_select":"URI"}},{"subitem_relation_name":[{"subitem_relation_name_text":"https://kaken.nii.ac.jp/ja/grant/KAKENHI-PROJECT-15790147/"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/ja/grant/KAKENHI-PROJECT-15790147/","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_creator":{"attribute_name":"著者","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{"creatorName":"置塩, 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