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Oligomerization of H+-pyrophosphatase and its structural and functional consequences
http://hdl.handle.net/2237/6661
4bf69973-9e69-48f4-b0c3-9e9e7f448d82
名前 / ファイル | ライセンス | アクション | |
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Item type | 学術雑誌論文 / Journal Article(1) | |||||
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公開日 | 2006-06-30 | |||||
タイトル | ||||||
タイトル | Oligomerization of H+-pyrophosphatase and its structural and functional consequences | |||||
著者 |
Mimura, Hisatoshi
× Mimura, Hisatoshi× Nakanishi, Yoichi× Maeshima, Masayoshi× 前島, 正義 |
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キーワード | ||||||
主題Scheme | Other | |||||
主題 | Disulfide bond | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | H+-pyrophosphatase | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Oligomerization | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Proton pump | |||||
抄録 | ||||||
内容記述 | The H+-pyrophosphatase (H+-PPase) consists of a single polypeptide, containing 16 or 17 transmembrane domains. To determine the higher order oligomeric state of Streptomyces coelicolor H+-PPase, we constructed a series of cysteine substitution mutants and expressed them in Escherichia coli. Firstly, we analyzed the formation of disulfide bonds, promoted by copper, in mutants with single cysteine substitutions. 28 of 39 mutants formed disulfide bonds, including S545C, a substitution at the periplasmic side. The formation of intermolecular disulfide bonds suppressed the enzyme activity of several, where the substituted residues were located in the cytosol. Creating disulfide links in the cytosol may interfere with the enzyme’s catalytic function. Secondly, we prepared double mutants by introducing second cysteine substitutions into the S545C mutant. These double-cysteine mutants produced cross-linked complexes, estimated to be at least tetramers and possibly hexamers. Thirdly, we co-expressed epitope-tagged, wild type and inactive mutant H+-PPases in E. coli and confirmed the formation of oligomers by co-purifying one subunit using the epitope tag used to label the other. The enzyme activity of these oligomers was markedly suppressed. We propose that H+-PPase is present as an oligomer made up of at least two or three sets of dimers. |
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内容記述タイプ | Abstract | |||||
出版者 | ||||||
出版者 | Elsevier | |||||
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 | |||||
書誌情報 |
Biochimica et Biophysica Acta 巻 1708, 号 3, p. 393-403, 発行日 2005-07 |
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フォーマット | ||||||
application/pdf | ||||||
著者版フラグ | ||||||
値 | publisher | |||||
URI | ||||||
識別子 | http://hdl.handle.net/2237/6661 | |||||
識別子タイプ | HDL |