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Membrane Topology of the H+-Pyrophosphatase of Streptomycescoelicolor Determined by Cysteine-Scanning Mutagenesis
http://hdl.handle.net/2237/7089
http://hdl.handle.net/2237/70890e921ee6-6448-440a-9b5f-4f63f934bf9f
| 名前 / ファイル | ライセンス | アクション |
|---|---|---|
2004-JBC-ScPP.pdf (965.5 kB)
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| Item type | 学術雑誌論文 / Journal Article(1) | |||||
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| 公開日 | 2006-10-27 | |||||
| タイトル | ||||||
| タイトル | Membrane Topology of the H+-Pyrophosphatase of Streptomycescoelicolor Determined by Cysteine-Scanning Mutagenesis | |||||
| 言語 | en | |||||
| 著者 |
Mimura, Hisatoshi
× Mimura, Hisatoshi× Nakanishi, Yoichi× Hirono, Megumi× Maeshima, Masayoshi |
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| アクセス権 | ||||||
| アクセス権 | open access | |||||
| アクセス権URI | http://purl.org/coar/access_right/c_abf2 | |||||
| 抄録 | ||||||
| 内容記述タイプ | Abstract | |||||
| 内容記述 | 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. | |||||
| 言語 | en | |||||
| 出版者 | ||||||
| 出版者 | the American Society for Biochemistry and Molecular Biology | |||||
| 言語 | en | |||||
| 言語 | ||||||
| 言語 | eng | |||||
| 資源タイプ | ||||||
| 資源タイプresource | http://purl.org/coar/resource_type/c_6501 | |||||
| タイプ | journal article | |||||
| 出版タイプ | ||||||
| 出版タイプ | VoR | |||||
| 出版タイプResource | http://purl.org/coar/version/c_970fb48d4fbd8a85 | |||||
| DOI | ||||||
| 関連タイプ | isVersionOf | |||||
| 識別子タイプ | DOI | |||||
| 関連識別子 | https://doi.org/10.1074/jbc.M406264200 | |||||
| ISSN | ||||||
| 収録物識別子タイプ | PISSN | |||||
| 収録物識別子 | 0021-9258 | |||||
| 書誌情報 |
en : Journal of Biological chemistry 巻 279, p. 35106-35112, 発行日 2004-08 |
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| フォーマット | ||||||
| 値 | application/pdf | |||||
| 著者版フラグ | ||||||
| 値 | publisher | |||||
| URI | ||||||
| 識別子 | http://hdl.handle.net/2237/7089 | |||||
| 識別子タイプ | HDL | |||||
