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Isolation, characterization and electron microscopic single particle analysis of the NADH : ubiquinone oxidoreductase (complex I) from the hyperthermophilic eubacterium Aquifex aeolicus | |
Peng, GH; Fritzsch, G; Zickermann, V; Schaagger, H; Mentele, R; Lottspeich, F; Bostina, M; Radermacher, M; Huber, R; Stetter, KO; Michel, H; Michel, H, Max Planck Inst Biophys, Frankfurt, Germany | |
2003-03-18 | |
发表期刊 | BIOCHEMISTRY |
ISSN | 0006-2960 |
卷号 | 42期号:10页码:3032-3039 |
文章类型 | Article |
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. |
关键词 | Escherichia-coli Mitochondrial Nadh Bacterial Protein Purification Translocation Dehydrogenase Flexibility Inhibitors Stability |
学科领域 | Biochemistry & Molecular Biology |
DOI | 10.1021/bi026876v |
URL | 查看原文 |
收录类别 | SCI |
语种 | 英语 |
WOS记录号 | WOS:000181535300030 |
引用统计 | |
文献类型 | 期刊论文 |
条目标识符 | http://ir.qdio.ac.cn/handle/337002/1659 |
专题 | 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. |
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