| ○種別 (必須): | □ | 学術論文 (審査論文)
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| ○言語 (必須): | □ | 英語
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| ○著者 (必須): | 1. | (英) Yamagoshi Ryohei (日) (読)
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| 2. | 山本 武範
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| 3. | 橋本 満 ([熊本大学])
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| 4. | (英) Sugahara Ryohei (日) (読)
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| 5. | (英) Shiotsuki Takahiro (日) (読)
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| 6. | (英) Miyoshi Hideto (日) (読)
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| 7. | 寺田 弘 ([東京理科大学])
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| 8. | 篠原 康雄 ([徳島大学.先端酵素学研究所.基幹研究部門])
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| ○題名 (必須): | □ | (英) Identification of amino acid residues of mammalian mitochondrial phosphate carrier important for its functional expression in yeast cells, as achieved by PCR-mediated random mutation and gap-repair cloning. (日)
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| ○要約 (任意): | □ | (英) The mitochondrial phosphate carrier (PiC) of mammals, but not the yeast one, is synthesized with a presequence. The deletion of this presequence of the mammalian PiC was reported to facilitate the import of the carrier into yeast mitochondria, but the question as to whether or not mammalian PiC could be functionally expressed in yeast mitochondria was not addressed. In the present study, we first examined whether the defective growth on a glycerol plate of yeast cells lacking the yeast PiC gene could be reversed by the introduction of expression vectors of rat PiCs. The introduction of expression vectors encoding full-length rat PiC (rPiC) or rPiC lacking the presequence (NrPiC) was ineffective in restoring growth on the glycerol plates. When we examined the expression levels of individual rPiCs in yeast mitochondria, NrPiC was expressed at a level similar to that of yeast PiC, but that of rPiC was very low. These results indicated that NrPiC expressed in yeast mitochondria is inert. Next, we sought to isolate "revertants" viable on the glycerol plate by expressing randomly mutated NrPiC, and obtained two clones. These clones carried either of two mutations, F267S or F282S; and these mutations restored the transport function of NrPiC in yeast mitochondria. These two Phe residues were conserved in human carrier (hPiC), and the transport function of NhPiC expressed in yeast mitochondria was also markedly improved by their substitutions. Thus, substitution of F267S or F282S was concluded to be important for functional expression of mammalian PiCs in yeast mitochondria. (日)
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| ○キーワード (推奨): |
| ○発行所 (推奨): |
| ○誌名 (必須): | □ | Mitochondrion (Mitochondria Research Society)
(pISSN: 1567-7249, eISSN: 1872-8278)
| ○ISSN (任意): | □ | 1872-8278
ISSN: 1567-7249
(pISSN: 1567-7249, eISSN: 1872-8278) Title: MitochondrionTitle(ISO): MitochondrionPublisher: Elsevier B.V. (NLM Catalog)
(Scopus)
(CrossRef)
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| ○巻 (必須): | □ | 32
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| ○号 (必須): | □ |
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| ○頁 (必須): | □ | 1 9
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| ○年月日 (必須): | □ | 西暦 2016年 11月 9日 (平成 28年 11月 9日)
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| ○URL (任意): |
| ○DOI (任意): | □ | 10.1016/j.mito.2016.11.003 (→Scopusで検索)
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| ○PMID (任意): | □ | 27836624 (→Scopusで検索)
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| ○備考 (任意): | 1. | (英) PublicationType: Journal Article (日)
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