A role for native lipids in the stabilization and two dimensional crystallization of the Escherichia coli NADH ubiquinone oxidoreductase (complex I)

L.A. Sazanov, J. Carroll, P. Holt, L. Toime, I. Fearnley, Journal of Biological Chemistry 278 (2003) 19483–19491.

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Journal Article | Published
Author
Sazanov, Leonid AIST Austria ; Carroll, Joe D; Holt, Peter J; Toime, Laurence J; Fearnley, Ian M
Abstract
NADH-ubiquinone oxidoreductase (complex I or NDH-1) was purified from the BL21 strain of Escherichia coli using an improved procedure. The complex was effectively stabilized by addition of divalent cations and lipids, making the preparation suitable for structural studies. The ubiquinone reductase activity of the enzyme was fully restored by addition of native E. coli lipids. Two different two-dimensional crystal forms, with p2 and p3 symmetry, were obtained using lipids containing native E. coli extracts. Analysis of the crystals showed that they are formed by fully intact complex I in an L-shaped conformation. Activity assays and single particle analysis indicated that complex I maintains this structure in detergent solution and does not adopt a different conformation in the active state. Thus, we provide the first experimental evidence that complex I from E. coli has an L-shape in a lipid bilayer and confirm that this is also the case for the active enzyme in solution. This suggests strongly that bacterial complex I exists in an L-shaped conformation in vivo. Our results also indicate that native lipids play an important role in the activation, stabilization and, as a consequence, crystallization of purified complex I from E. coli.
Publishing Year
Date Published
2003-05-23
Journal Title
Journal of Biological Chemistry
Volume
278
Issue
21
Page
19483 - 19491
IST-REx-ID

Cite this

Sazanov LA, Carroll J, Holt P, Toime L, Fearnley I. A role for native lipids in the stabilization and two dimensional crystallization of the Escherichia coli NADH ubiquinone oxidoreductase (complex I). Journal of Biological Chemistry. 2003;278(21):19483-19491. doi:10.1074/jbc.M208959200
Sazanov, L. A., Carroll, J., Holt, P., Toime, L., & Fearnley, I. (2003). A role for native lipids in the stabilization and two dimensional crystallization of the Escherichia coli NADH ubiquinone oxidoreductase (complex I). Journal of Biological Chemistry, 278(21), 19483–19491. https://doi.org/10.1074/jbc.M208959200
Sazanov, Leonid A, Joe Carroll, Peter Holt, Laurence Toime, and Ian Fearnley. “A Role for Native Lipids in the Stabilization and Two Dimensional Crystallization of the Escherichia Coli NADH Ubiquinone Oxidoreductase (Complex I).” Journal of Biological Chemistry 278, no. 21 (2003): 19483–91. https://doi.org/10.1074/jbc.M208959200.
L. A. Sazanov, J. Carroll, P. Holt, L. Toime, and I. Fearnley, “A role for native lipids in the stabilization and two dimensional crystallization of the Escherichia coli NADH ubiquinone oxidoreductase (complex I),” Journal of Biological Chemistry, vol. 278, no. 21, pp. 19483–19491, 2003.
Sazanov LA, Carroll J, Holt P, Toime L, Fearnley I. 2003. A role for native lipids in the stabilization and two dimensional crystallization of the Escherichia coli NADH ubiquinone oxidoreductase (complex I). Journal of Biological Chemistry. 278(21), 19483–19491.
Sazanov, Leonid A., et al. “A Role for Native Lipids in the Stabilization and Two Dimensional Crystallization of the Escherichia Coli NADH Ubiquinone Oxidoreductase (Complex I).” Journal of Biological Chemistry, vol. 278, no. 21, American Society for Biochemistry and Molecular Biology, 2003, pp. 19483–91, doi:10.1074/jbc.M208959200.

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