Isolation, characterization and electron microscopic single particle analysis of the NADH : ubiquinone oxidoreductase (complex I) from the hyperthermophilic eubacterium Aquifex aeolicus
文献类型：期刊论文
| 作者 | Peng, GH; Fritzsch, G; Zickermann, V; Schaagger, H; Mentele, R; Lottspeich, F; Bostina, M; Radermacher, M; Huber, R; Stetter, KO |
| 刊名 | BIOCHEMISTRY
 |
| 出版日期 | 2003-03-18 |
| 卷号 | 42期号:10页码:3032-3039 |
| 关键词 | Escherichia-coli Mitochondrial Nadh Bacterial Protein Purification Translocation Dehydrogenase Flexibility Inhibitors Stability |
| ISSN号 | 0006-2960 |
| DOI | 10.1021/bi026876v |
| 文献子类 | Article |
| 英文摘要 | The proton-translocating NADH:ubiquinone oxidoreductase (complex I) has been purified from Aquifex aeolicus, a hyperthermophilic eubacterium of known genome sequence. The purified detergent solubilized enzyme is highly active above 50 degreesC. The specific activity for electron transfer from NADH to decylubiquinone is 29 U/mg at 80 degreesC. The A. aeolicus complex I is completely sensitive to rotenone and 2-n-decyl-quinazoline-4-yl-amine. SDS polyacrylamide gel electrophoresis shows that it may contain up to 14 subunits. N-terminal amino acid sequencing of the bands indicates the presence of a stable subcomplex, which is composed of subunits E, F, and G. The isolated complex is highly stable and active in a temperature range from 50 to 90 degreesC, with a half-life of about 10 h at 80 degreesC. The activity shows a linear Arrhenius plot at 50-85 degreesC with an activation energy at 31.92 J/mol K. Single particle electron microscopy shows that the A. aeolicus complex I has the typical L-shape. However, visual inspection of averaged images reveals many more details in the external arm of the complex than has been observed for complex I from other sources. In addition, the angle (90degrees) between the cytoplasmic peripheral arm and the membrane intrinsic arm of the complex appears to be invariant.; The proton-translocating NADH:ubiquinone oxidoreductase (complex I) has been purified from Aquifex aeolicus, a hyperthermophilic eubacterium of known genome sequence. The purified detergent solubilized enzyme is highly active above 50 degreesC. The specific activity for electron transfer from NADH to decylubiquinone is 29 U/mg at 80 degreesC. The A. aeolicus complex I is completely sensitive to rotenone and 2-n-decyl-quinazoline-4-yl-amine. SDS polyacrylamide gel electrophoresis shows that it may contain up to 14 subunits. N-terminal amino acid sequencing of the bands indicates the presence of a stable subcomplex, which is composed of subunits E, F, and G. The isolated complex is highly stable and active in a temperature range from 50 to 90 degreesC, with a half-life of about 10 h at 80 degreesC. The activity shows a linear Arrhenius plot at 50-85 degreesC with an activation energy at 31.92 J/mol K. Single particle electron microscopy shows that the A. aeolicus complex I has the typical L-shape. However, visual inspection of averaged images reveals many more details in the external arm of the complex than has been observed for complex I from other sources. In addition, the angle (90degrees) between the cytoplasmic peripheral arm and the membrane intrinsic arm of the complex appears to be invariant. |
| 学科主题 | Biochemistry & Molecular Biology |
| URL标识 | 查看原文 |
| 语种 | 英语 |
| WOS记录号 | WOS:000181535300030 |
| 公开日期 | 2010-11-18 |
| 源URL | [http://ir.qdio.ac.cn/handle/337002/1660]  |
| 专题 | 海洋研究所_实验海洋生物学重点实验室 |
| 通讯作者 | Michel, H, Max Planck Inst Biophys, Frankfurt, Germany |
| 作者单位 | 1.Max Planck Inst Biophys, Frankfurt, Germany 2.Chinese Acad Sci, Inst Oceanol, Qingdao, Peoples R China 3.Univ Frankfurt Klinikum, Gustav Embden Zentrum Biol Chem, D-6000 Frankfurt, Germany 4.Max Planck Inst Biochem, D-82152 Martinsried, Germany 5.Univ Regensburg, Lehrstuhl Mikrobiol, D-8400 Regensburg, Germany 6.Univ Regensburg, Archaeenzentrum, D-8400 Regensburg, Germany |
| 推荐引用方式 GB/T 7714 | Peng, GH,Fritzsch, G,Zickermann, V,et al. Isolation, characterization and electron microscopic single particle analysis of the NADH : ubiquinone oxidoreductase (complex I) from the hyperthermophilic eubacterium Aquifex aeolicus[J]. BIOCHEMISTRY,2003,42(10):3032-3039. |
| APA | Peng, GH.,Fritzsch, G.,Zickermann, V.,Schaagger, H.,Mentele, R.,...&Michel, H, Max Planck Inst Biophys, Frankfurt, Germany.(2003).Isolation, characterization and electron microscopic single particle analysis of the NADH : ubiquinone oxidoreductase (complex I) from the hyperthermophilic eubacterium Aquifex aeolicus.BIOCHEMISTRY,42(10),3032-3039. |
| MLA | Peng, GH,et al."Isolation, characterization and electron microscopic single particle analysis of the NADH : ubiquinone oxidoreductase (complex I) from the hyperthermophilic eubacterium Aquifex aeolicus".BIOCHEMISTRY 42.10(2003):3032-3039. |
入库方式： OAI收割
来源：海洋研究所
浏览0
下载0
收藏0
其他版本
除非特别说明，本系统中所有内容都受版权保护，并保留所有权利。