{"created":"2023-06-20T16:39:59.258279+00:00","id":10141,"links":{},"metadata":{"_buckets":{"deposit":"77650b7e-48b7-4f6c-b26b-0f67ef77f2d9"},"_deposit":{"created_by":3,"id":"10141","owners":[3],"pid":{"revision_id":0,"type":"depid","value":"10141"},"status":"published"},"_oai":{"id":"oai:kindai.repo.nii.ac.jp:00010141","sets":["14:923:2219:2220","21:3039:3435:3455"]},"author_link":["18977","18976","18974","18975"],"item_2_alternative_title_3":{"attribute_name":"その他（別言語等）のタイトル","attribute_value_mlt":[{"subitem_alternative_title":"メタノール資化酵母を用いた蛋白質異常凝集解離新規シャペロンの発現系の検討"}]},"item_2_biblio_info_21":{"attribute_name":"書誌情報","attribute_value_mlt":[{"bibliographicIssueDates":{"bibliographicIssueDate":"2015-03-01","bibliographicIssueDateType":"Issued"},"bibliographicPageEnd":"41","bibliographicPageStart":"31","bibliographicVolumeNumber":"20","bibliographic_titles":[{"bibliographic_title":"近畿大学先端技術総合研究所紀要"},{"bibliographic_title":"Memoirs of Institute of Advanced Technology, Kinki University","bibliographic_titleLang":"en"}]}]},"item_2_description_33":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"[Abstract] Oligomeric state of the actin interacting protein 2/D-lactate dehydrogenase protein 2（ Aip2p/Dld2p）isolated from Saccharomyces cerevisiae was shown to be able to unfold the conformation of pathogenic highly aggregated polypeptides, and has been supposed to be a novel chaperone protein. However, the mechanism of the unfolding activity is little understood. Since experiments for the structure–function relationships of a protein require an efficient expression system, we attempted to use the methylotrophic yeast Pichia pastoris because of its ability of very high expression yield. Changing the vector/protein construct combinations, we examined the secretion expression system of P. pastoris with several Aip2p/Dld2p constructs as well as the intracellular expression system. We did not observe significant expression yields in any construct combinations. RT-PCR analyses revealed that mRNA coding Aip2p/Dld2p was successfully synthesized, suggesting that the structure of Aip2p/Dld2p is inherently unstable and susceptible to proteolysis of the cellular quality control system.                    [抄録]出芽酵母Saccharomyces cerevisiae から単離されたオリゴメリック蛋白質Aip2p/Dld2p は病原性の蛋白質異常凝集体を解きほぐす活性を持ち, 新規のシャペロン蛋白質であることが示唆されている. しかしながら, その解きほぐし活性の物理化学的メカニズムについてはほとんど調べられていない. 蛋白質の物理化学的解析や構造解析には数十ミリグラム程度の試料が必要であり, そのためには高効率の蛋白質発現系を要する. そこで今回, 我々は他の蛋白質で高収量が報告されているメタノール資化酵母Pichia pastorisによる大量発現を試みた. 種々のAip2p/Dld2p コンストラクト（全長体、N 末端欠損体, システイン→アラニン置換体）とベクター（細胞内発現系、分泌発現系）の組み合わせを試したところ, Aip2p/Dld2p をコードするmRNA の合成は見られたが, 蛋白質の発現は細胞内発現系でも分泌発現系でも見られず, さらに分泌発現系の場合の細胞内にも見られなかった. Aip2p/Dld2p の天然構造は非常に不安定で, ヘテロロガスな発現の場合, 翻訳直後に細胞内の品質管理機構によって加水分解を受けているのではないかと考えられる.","subitem_description_type":"Abstract"}]},"item_2_description_36":{"attribute_name":"内容記述","attribute_value_mlt":[{"subitem_description":"近畿大学先端技術総合研究所紀要編集委員会","subitem_description_type":"Other"}]},"item_2_description_41":{"attribute_name":"フォーマット","attribute_value_mlt":[{"subitem_description":"application/pdf","subitem_description_type":"Other"}]},"item_2_publisher_14":{"attribute_name":"出版者 名前","attribute_value_mlt":[{"subitem_publisher":"近畿大学先端技術総合研究所"}]},"item_2_source_id_22":{"attribute_name":"ISSN","attribute_value_mlt":[{"subitem_source_identifier":"13468693","subitem_source_identifier_type":"ISSN"}]},"item_2_text_7":{"attribute_name":"著者(英)","attribute_value_mlt":[{"subitem_text_language":"en","subitem_text_value":"Sakurai, Kazumasa"},{"subitem_text_language":"en","subitem_text_value":"Nakata, Shoki"},{"subitem_text_language":"en","subitem_text_value":"Toyomasu, Akihiro"},{"subitem_text_language":"en","subitem_text_value":"Tachibana, Hideki"}]},"item_2_text_8":{"attribute_name":"著者 所属","attribute_value_mlt":[{"subitem_text_value":"High Pressure Protein Research Center, Institute of Advanced Technology, Kinki University"},{"subitem_text_value":"Department of Biotechnology, Kinki University"},{"subitem_text_value":"Department of Biotechnology, Kinki University"},{"subitem_text_value":"High Pressure Protein Research Center, Institute of Advanced Technology, Kinki University : Department of Biotechnology, Kinki University"}]},"item_2_text_9":{"attribute_name":"著者所属(翻訳)","attribute_value_mlt":[{"subitem_text_value":"Kinki University"},{"subitem_text_value":"Kinki University"},{"subitem_text_value":"Kinki University"},{"subitem_text_value":"Kinki University"}]},"item_2_version_type_12":{"attribute_name":"版","attribute_value_mlt":[{"subitem_version_resource":"http://purl.org/coar/version/c_970fb48d4fbd8a85","subitem_version_type":"VoR"}]},"item_creator":{"attribute_name":"著者","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{"creatorName":"櫻井,  一正"},{"creatorName":"サクライ, カズマサ","creatorNameLang":"ja-Kana"}],"nameIdentifiers":[{}]},{"creatorNames":[{"creatorName":"中田,  翔貴"},{"creatorName":"ナカタ, ショウキ","creatorNameLang":"ja-Kana"}],"nameIdentifiers":[{}]},{"creatorNames":[{"creatorName":"豊増,  明博"},{"creatorName":"トヨマス, アキヒロ","creatorNameLang":"ja-Kana"}],"nameIdentifiers":[{}]},{"creatorNames":[{"creatorName":"橘,  秀樹"},{"creatorName":"タチバナ, ヒデキ","creatorNameLang":"ja-Kana"}],"nameIdentifiers":[{}]}]},"item_files":{"attribute_name":"ファイル情報","attribute_type":"file","attribute_value_mlt":[{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2016-02-18"}],"displaytype":"detail","filename":"AA11574630-20150331-0031.pdf","filesize":[{"value":"851.5 kB"}],"format":"application/pdf","licensetype":"license_note","mimetype":"application/pdf","url":{"label":"AA11574630-20150331-0031.pdf","url":"https://kindai.repo.nii.ac.jp/record/10141/files/AA11574630-20150331-0031.pdf"},"version_id":"c285b26b-f912-4ba5-9810-e0d83c25044c"}]},"item_keyword":{"attribute_name":"キーワード","attribute_value_mlt":[{"subitem_subject":"Aip2p/Dld2p","subitem_subject_scheme":"Other"},{"subitem_subject":"Pichia pastoris","subitem_subject_scheme":"Other"},{"subitem_subject":"protein aggregation","subitem_subject_scheme":"Other"},{"subitem_subject":"unfolding activity","subitem_subject_scheme":"Other"},{"subitem_subject":"Aip2p/Dld2p","subitem_subject_scheme":"Other"},{"subitem_subject":"メタノール資化酵母Pichia pastoris","subitem_subject_scheme":"Other"},{"subitem_subject":"異常凝集体","subitem_subject_scheme":"Other"},{"subitem_subject":"解きほぐし活性","subitem_subject_scheme":"Other"}]},"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":"departmental bulletin paper","resourceuri":"http://purl.org/coar/resource_type/c_6501"}]},"item_title":"<研究ノート>EXAMINATION OF THE EXPRESSION OF NOVEL CHAPERONE PROTEIN AIP2P/DLD2P WITH METHYLOTROPHIC YEAST PICHIA PASTORIS","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"<研究ノート>EXAMINATION OF THE EXPRESSION OF NOVEL CHAPERONE PROTEIN AIP2P/DLD2P WITH METHYLOTROPHIC YEAST PICHIA PASTORIS"}]},"item_type_id":"2","owner":"3","path":["2220","3455"],"pubdate":{"attribute_name":"公開日","attribute_value":"2015-05-29"},"publish_date":"2015-05-29","publish_status":"0","recid":"10141","relation_version_is_last":true,"title":["<研究ノート>EXAMINATION OF THE EXPRESSION OF NOVEL CHAPERONE PROTEIN AIP2P/DLD2P WITH METHYLOTROPHIC YEAST PICHIA PASTORIS"],"weko_creator_id":"3","weko_shared_id":-1},"updated":"2023-06-20T23:46:09.678541+00:00"}