{"created":"2023-06-20T16:46:12.528425+00:00","id":17645,"links":{},"metadata":{"_buckets":{"deposit":"d9393643-c694-46d4-876b-f9a3277c2cc0"},"_deposit":{"created_by":29,"id":"17645","owners":[29],"pid":{"revision_id":0,"type":"depid","value":"17645"},"status":"published"},"_oai":{"id":"oai:kindai.repo.nii.ac.jp:00017645","sets":["14:923:2219:4273"]},"author_link":["29824","29823"],"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":"大腸菌によるproMTG 発現精製と圧力変性"}]},"item_2_biblio_info_21":{"attribute_name":"書誌情報","attribute_value_mlt":[{"bibliographicIssueDates":{"bibliographicIssueDate":"2016-05-31","bibliographicIssueDateType":"Issued"},"bibliographicPageEnd":"22","bibliographicPageStart":"11","bibliographicVolumeNumber":"21","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, Kindai University","bibliographic_titleLang":"en"}]}]},"item_2_description_33":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"Understanding of pressure effect on conformational change of proteins will allow a method by which enzymatic activities are freely manipulated by pressure application. For this goal, we investigated the pressure effect on the proenzyme of microbial transglutaminase（ MTG）. First, we examined the E. coli protein expression system. Then, we found an effective expression conditions. Subsequently, we measured the pressure-induced unfolding of the obtained proMTG. The results indicate that the native conformation of proMTG has a relatively low volume, indicating that structural rigidity is enhanced in the proenzyme state. We suppose that the enhanced rigidity likely contribute to the suppression of its enzymatic activity.\n酵素活性は加圧条件下で変化することが知られている。圧力のこの効果の分子的なメカニズムを理解し加圧による酵素機能の制御が可能になれば、幅広い応用が予想される。そこで、我々は加圧による微生物由来トランスグルタミナーゼ（ MTG）の構造への影響を調べることを目指した。タンパク質架橋化酵素、トランスグルタミナーゼ（TG）は幅広く生物界に存在し、基質タンパク質どうしを架橋する機能を持つ酵素である。はじめに我々はMTG のプロ酵素であるproMTG の大腸菌による大量発現の系の確立を目指した。発現条件検討の結果、20℃で長時間の発現誘導を行うと、正しい構造を持った状態で発現することが分かり、効率の良いproMTG 発現条件を決定できた。得られたproMTG を用いて圧力蛍光測定実験を行ったところ、圧力感受性が非常に低いことが分かった。これは変性に伴う体積変化（ΔV）の絶対値が小さいことを示す。一般的に変性に伴い、蛋白質分子のモル体積は減少し、天然状態の分子構造内にボイドと呼ばれる隙間が多いとΔV の絶対値は大きくなるが、proMTG ではプロ配列が酵素の活性部位にクレフトにぴったり挟まっており、これが分子構造に隙間のない状態にしているため低いΔV が得られたと考えられる。構造が密になることでpro 状態の分子の剛直性を上げ、その結果酵素活性が抑制されているものと考えられる。","subitem_description_type":"Abstract"}]},"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":"Takashiba, Kazuma"},{"subitem_text_language":"en","subitem_text_value":"Sakurai, Kazumasa"}]},"item_2_text_8":{"attribute_name":"著者 所属","attribute_value_mlt":[{"subitem_text_value":"近畿大学生物理工学部生物工学科"},{"subitem_text_value":"近畿大学先端技術総合研究所高圧力蛋白質研究センター"}]},"item_2_text_9":{"attribute_name":"著者所属(翻訳)","attribute_value_mlt":[{"subitem_text_value":"Kindai University"},{"subitem_text_value":"Kindai 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":"髙芝, 一真"},{"creatorName":"タカシバ, カズマ","creatorNameLang":"ja-Kana"}],"nameIdentifiers":[{}]},{"creatorNames":[{"creatorName":"櫻井, 一正"},{"creatorName":"サクライ, カズマサ","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":"2016-08-02"}],"displaytype":"detail","filename":"AA11574630-20160531-0011.pdf","filesize":[{"value":"1.1 MB"}],"format":"application/pdf","licensetype":"license_note","mimetype":"application/pdf","url":{"label":"AA11574630-20160531-0011.pdf","url":"https://kindai.repo.nii.ac.jp/record/17645/files/AA11574630-20160531-0011.pdf"},"version_id":"954a4e16-a511-4024-bf49-17e3aba62870"}]},"item_keyword":{"attribute_name":"キーワード","attribute_value_mlt":[{"subitem_subject":"微生物トランスグルタミナーゼ","subitem_subject_scheme":"Other"},{"subitem_subject":"プロ酵素","subitem_subject_scheme":"Other"},{"subitem_subject":"蛋白質の圧力変性","subitem_subject_scheme":"Other"},{"subitem_subject":"microbial transglutaminase","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"proenzyme","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"pressure-induced unfolding","subitem_subject_language":"en","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":"〈Original Papers〉Expression of proenzyme of microbial transglutaminase and its pressure-dependent unfolding behavior","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"〈Original Papers〉Expression of proenzyme of microbial transglutaminase and its pressure-dependent unfolding behavior"}]},"item_type_id":"2","owner":"29","path":["4273"],"pubdate":{"attribute_name":"公開日","attribute_value":"2016-08-02"},"publish_date":"2016-08-02","publish_status":"0","recid":"17645","relation_version_is_last":true,"title":["〈Original Papers〉Expression of proenzyme of microbial transglutaminase and its pressure-dependent unfolding behavior"],"weko_creator_id":"29","weko_shared_id":-1},"updated":"2023-06-20T22:03:47.768802+00:00"}