| Item type |
文献 / Documents(1) |
| 公開日 |
2019-10-15 |
| アクセス権 |
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|
アクセス権 |
open access |
| 資源タイプ |
|
|
資源タイプ識別子 |
http://purl.org/coar/resource_type/c_6501 |
|
資源タイプ |
journal article |
| 出版社版DOI |
|
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|
識別子タイプ |
DOI |
|
|
関連識別子 |
https://doi.org/10.1016/j.mito.2016.11.003 |
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|
言語 |
ja |
|
|
関連名称 |
10.1016/j.mito.2016.11.003 |
| 出版タイプ |
|
|
出版タイプ |
AM |
|
出版タイプResource |
http://purl.org/coar/version/c_ab4af688f83e57aa |
| タイトル |
|
|
タイトル |
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 |
|
言語 |
en |
| 著者 |
ヤマゴシ, リョウヘイ
山本, 武範
橋本, 満
スガハラ, リョウヘイ
シオツキ, タカヒロ
ミヨシ, ヒデト
寺田, 弘
篠原, 康雄
|
| 抄録 |
|
|
内容記述タイプ |
Abstract |
|
内容記述 |
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 (ΔNrPiC) was ineffective in restoring growth on the glycerol plates. When we examined the expression levels of individual rPiCs in yeast mitochondria, ΔNrPiC was expressed at a level similar to that of yeast PiC, but that of rPiC was very low. These results indicated that ΔNrPiC expressed in yeast mitochondria is inert. Next, we sought to isolate “revertants” viable on the glycerol plate by expressing randomly mutated ΔNrPiC, and obtained two clones. These clones carried either of two mutations, F267S or F282S; and these mutations restored the transport function of ΔNrPiC in yeast mitochondria. These two Phe residues were conserved in human carrier (hPiC), and the transport function of ΔNhPiC 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. |
|
言語 |
en |
| キーワード |
|
|
言語 |
en |
|
主題Scheme |
Other |
|
主題 |
Mitochondrial solute carrier |
| キーワード |
|
|
言語 |
en |
|
主題Scheme |
Other |
|
主題 |
Phosphate carrier |
| キーワード |
|
|
言語 |
en |
|
主題Scheme |
Other |
|
主題 |
Functional expression |
| キーワード |
|
|
言語 |
en |
|
主題Scheme |
Other |
|
主題 |
Gap-repair cloning |
| キーワード |
|
|
言語 |
en |
|
主題Scheme |
Other |
|
主題 |
Yeast |
| 書誌情報 |
en : Mitochondrion
巻 32,
p. 1-9,
発行日 2016-11-09
|
| 収録物ID |
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|
収録物識別子タイプ |
ISSN |
|
収録物識別子 |
15677249 |
| 収録物ID |
|
|
収録物識別子タイプ |
NCID |
|
収録物識別子 |
AA11694235 |
| 収録物ID |
|
|
収録物識別子タイプ |
NCID |
|
収録物識別子 |
AA11643835 |
| 出版者 |
|
|
出版者 |
Elsevier |
|
言語 |
en |
| 出版者 |
|
|
出版者 |
Mitochondria Research Society |
|
言語 |
en |
| 権利情報 |
|
|
言語 |
en |
|
権利情報 |
© 2016. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/ |
| EID |
|
|
識別子 |
321434 |
|
識別子タイプ |
URI |
| 言語 |
|
|
言語 |
eng |
mito_32_1.pdf (775 KB)
