Isolation, characterization and electron microscopic single particle analysis of the NADH : ubiquinone oxidoreductase (complex I) from the hyperthermophilic eubacterium Aquifex aeolicus

doi:10.1021/bi026876v

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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	被引频次：73[WOS] [WOS记录] [WOS相关记录]
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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