{"created":"2021-03-01T06:12:02.847349+00:00","id":5483,"links":{},"metadata":{"_buckets":{"deposit":"fae13e38-b34b-4401-a991-df5f2c12a173"},"_deposit":{"id":"5483","owners":[],"pid":{"revision_id":0,"type":"depid","value":"5483"},"status":"published"},"_oai":{"id":"oai:nagoya.repo.nii.ac.jp:00005483","sets":["643:666:667"]},"author_link":["14144","14145","14146","14147"],"item_10_biblio_info_6":{"attribute_name":"書誌情報","attribute_value_mlt":[{"bibliographicIssueDates":{"bibliographicIssueDate":"2004-08","bibliographicIssueDateType":"Issued"},"bibliographicPageEnd":"35112","bibliographicPageStart":"35106","bibliographicVolumeNumber":"279","bibliographic_titles":[{"bibliographic_title":"Journal of Biological chemistry","bibliographic_titleLang":"en"}]}]},"item_10_description_4":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"The H^+-translocating pyrophosphatase (H^+-PPase) is a proton pump that is found in a wide variety of organisms. It consists of a single polypeptide chain that is thought to possess between 14 and 17 transmembrane domains. To determine the topological arrangement of its conserved motifs and transmembrane domains, we carried out a cysteine-scanning analysis by determining the membrane topology of cysteine-substitution mutants of Streptomyces coelicolor H^+-PPase expressed in Escherichia coli, using chemical reagents. First, we prepared a synthetic DNA that encoded the enzyme and constructed a functional cysteine-less mutant by substituting the four cysteine residues. We then introduced cysteine residues individually into 42 sites in its hydrophilic regions, and N- and C-terminal segments. Thirtysix of the mutant enzymes retained both pyrophosphatase and H^+-translocating activities. Analysis of 29 of these mutant forms using membrane permeable and impermeable sulfhydryl reagents revealed that S. coelicolor H^+-PPase contains 17 transmembrane domains, and that several conserved segments, such as the substrate-binding domains, are exposed to the cytoplasm. Four essential serine residues that were located on the cytoplasmic side were also identified. A marked characteristic of the S. coelicolor enzyme is a long additional sequence, which includes a transmembrane domain at the C-terminus. We propose that the basic structure of H^+-PPases has 16 transmembrane domains with several large cytoplasmic loops containing functional motifs.","subitem_description_language":"en","subitem_description_type":"Abstract"}]},"item_10_identifier_60":{"attribute_name":"URI","attribute_value_mlt":[{"subitem_identifier_type":"HDL","subitem_identifier_uri":"http://hdl.handle.net/2237/7089"}]},"item_10_publisher_32":{"attribute_name":"出版者","attribute_value_mlt":[{"subitem_publisher":"the American Society for Biochemistry and Molecular Biology","subitem_publisher_language":"en"}]},"item_10_relation_11":{"attribute_name":"DOI","attribute_value_mlt":[{"subitem_relation_type":"isVersionOf","subitem_relation_type_id":{"subitem_relation_type_id_text":"https://doi.org/10.1074/jbc.M406264200","subitem_relation_type_select":"DOI"}}]},"item_10_select_15":{"attribute_name":"著者版フラグ","attribute_value_mlt":[{"subitem_select_item":"publisher"}]},"item_10_source_id_7":{"attribute_name":"ISSN","attribute_value_mlt":[{"subitem_source_identifier":"0021-9258","subitem_source_identifier_type":"PISSN"}]},"item_10_text_14":{"attribute_name":"フォーマット","attribute_value_mlt":[{"subitem_text_value":"application/pdf"}]},"item_1615787544753":{"attribute_name":"出版タイプ","attribute_value_mlt":[{"subitem_version_resource":"http://purl.org/coar/version/c_970fb48d4fbd8a85","subitem_version_type":"VoR"}]},"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":"Mimura, Hisatoshi","creatorNameLang":"en"}],"nameIdentifiers":[{"nameIdentifier":"14144","nameIdentifierScheme":"WEKO"}]},{"creatorNames":[{"creatorName":"Nakanishi, Yoichi","creatorNameLang":"en"}],"nameIdentifiers":[{"nameIdentifier":"14145","nameIdentifierScheme":"WEKO"}]},{"creatorNames":[{"creatorName":"Hirono, Megumi","creatorNameLang":"en"}],"nameIdentifiers":[{"nameIdentifier":"14146","nameIdentifierScheme":"WEKO"}]},{"creatorNames":[{"creatorName":"Maeshima, Masayoshi","creatorNameLang":"en"}],"nameIdentifiers":[{"nameIdentifier":"14147","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":"2018-02-19"}],"displaytype":"detail","filename":"2004-JBC-ScPP.pdf","filesize":[{"value":"965.5 kB"}],"format":"application/pdf","licensetype":"license_note","mimetype":"application/pdf","url":{"label":"2004-JBC-ScPP.pdf","objectType":"fulltext","url":"https://nagoya.repo.nii.ac.jp/record/5483/files/2004-JBC-ScPP.pdf"},"version_id":"6283b91f-8b41-4a16-8c50-e76ca4a2a67d"}]},"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":"journal article","resourceuri":"http://purl.org/coar/resource_type/c_6501"}]},"item_title":"Membrane Topology of the H+-Pyrophosphatase of Streptomycescoelicolor Determined by Cysteine-Scanning Mutagenesis","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"Membrane Topology of the H+-Pyrophosphatase of Streptomycescoelicolor Determined by Cysteine-Scanning Mutagenesis","subitem_title_language":"en"}]},"item_type_id":"10","owner":"1","path":["667"],"pubdate":{"attribute_name":"PubDate","attribute_value":"2006-10-27"},"publish_date":"2006-10-27","publish_status":"0","recid":"5483","relation_version_is_last":true,"title":["Membrane Topology of the H+-Pyrophosphatase of Streptomycescoelicolor Determined by Cysteine-Scanning Mutagenesis"],"weko_creator_id":"1","weko_shared_id":-1},"updated":"2023-01-16T03:54:22.091856+00:00"}