CDNA cloning, functional expression and cellular localization of rat liver mitochondrial electron-transfer flavoprotein-ubiquinone oxidoreductase protein
文献类型：期刊论文
| 作者 | Huang, SB; Wei, S; Lin, QH |
| 刊名 | SCIENCE IN CHINA SERIES C-LIFE SCIENCES
 |
| 出版日期 | 2005 |
| 卷号 | 48期号:4页码:357-367 |
| 关键词 | electron transfer flavoprotein ubiquinone oxidoreductase precursor rat protein purification gene clone tissue specificity |
| 通讯作者 | Wei, S (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Key Lab Mol Cell Biol, Inst Biochem & Cell Biol, Shanghai 200031, Peoples R China.,song_wei@sibs.ac.cn |
| 英文摘要 | A membrane-bound protein was purified from rat liver mitochondria. After being digested with V8 protease, two peptides containing identical 14 amino acid residue sequences were obtained. Using the 14 amino acid peptide derived DNA sequence as gene specific primer, the cDNA of correspondent gene F-terminal and 3'-terminal were obtained by RACE technique. The full-length cDNA that encoded a protein of 616 amino acids was thus cloned, which included the above mentioned peptide sequence. The full length cDNA was highly homologous to that of human ETF-QO, indicating that it may be the cDNA of rat ETF-QO. ETF-QO is an iron sulfur protein located in mitochondria inner membrane containing two kinds of redox center: FAD and [4Fe-4S] center. After comparing the sequence from the cDNA of the 616 amino acids protein with that of the mature protein of rat liver mitochondria, it was found that the N terminal 32 amino acid residues did not exist in the mature protein, indicating that the cDNA was that of ETF-QOp. When the cDNA was expressed in Saccharomyces cerevisiae with inducible vectors, the protein product was enriched in mitochondrial fraction and exhibited electron transfer activity (NBT reductase activity) of ETF-QO. Results demonstrated that the 32 amino acid peptide was a mitochondrial targeting peptide, and both FAD and iron-sulfur cluster were inserted properly into the expressed ETF-QO. ETF-QO had a high level expression in rat heart, liver and kidney. The fusion protein of GFP-ETF-QO co-localized with mitochondria in COS-7 cells. |
| 学科主题 | Life Sciences & Biomedicine - Other Topics |
| 类目[WOS] | Biology |
| 关键词[WOS] | COA DEHYDROGENASES ; RESPIRATORY-CHAIN ; PIG-LIVER ; ACYL-COA ; PURIFICATION ; IMPORT ; YEAST |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000232174800006 |
| 版本 | 出版稿 |
| 源URL | [http://202.127.25.143/handle/331003/1889]  |
| 专题 | 上海生化细胞研究所_上海生科院生化细胞研究所 |
| 推荐引用方式 GB/T 7714 | Huang, SB,Wei, S,Lin, QH. CDNA cloning, functional expression and cellular localization of rat liver mitochondrial electron-transfer flavoprotein-ubiquinone oxidoreductase protein[J]. SCIENCE IN CHINA SERIES C-LIFE SCIENCES,2005,48(4):357-367. |
| APA | Huang, SB,Wei, S,&Lin, QH.(2005).CDNA cloning, functional expression and cellular localization of rat liver mitochondrial electron-transfer flavoprotein-ubiquinone oxidoreductase protein.SCIENCE IN CHINA SERIES C-LIFE SCIENCES,48(4),357-367. |
| MLA | Huang, SB,et al."CDNA cloning, functional expression and cellular localization of rat liver mitochondrial electron-transfer flavoprotein-ubiquinone oxidoreductase protein".SCIENCE IN CHINA SERIES C-LIFE SCIENCES 48.4(2005):357-367. |
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