Abstract
Complex I is the first and largest enzyme of the respiratory chain, coupling electron transfer between NADH and ubiquinone to the translocation of four protons across the membrane. It has a central role in cellular energy production and has been implicated in many human neurodegenerative diseases. The L-shaped enzyme consists of hydrophilic and membrane domains. Previously, we determined the structure of the hydrophilic domain. Here we report the crystal structure of the Esherichia coli complex I membrane domain at 3.0 Å resolution. It includes six subunits, NuoL, NuoM, NuoN, NuoA, NuoJ and NuoK, with 55 transmembrane helices. The fold of the homologous antiporter-like subunits L, M and N is novel, with two inverted structural repeats of five transmembrane helices arranged, unusually, face-to-back. Each repeat includes a discontinuous transmembrane helix and forms half of a channel across the membrane. A network of conserved polar residues connects the two half-channels, completing the proton translocation pathway. Unexpectedly, lysines rather than carboxylate residues act as the main elements of the proton pump in these subunits. The fourth probable proton-translocation channel is at the interface of subunits N, K, J and A. The structure indicates that proton translocation in complex I, uniquely, involves coordinated conformational changes in six symmetrical structural elements.
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Acknowledgements
This work was funded by the Medical Research Council. We thank the ESRF and the SLS for provision of synchrotron radiation facilities. We are grateful to the staff of beamlines ID29 (ESRF) and X06SA (SLS) for assistance.
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R.G.E. performed research and analysed data; L.A.S. designed the project, analysed data and wrote the manuscript, with contributions from R.G.E.
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Supplementary Information
This file contains a Supplementary Discussion, Supplementary Tables 1-8, Supplementary Figures 1-8 with legends and additional references. (PDF 7855 kb)
Supplementary Movie 1
This movie illustrates the overall architecture of the membrane domain, showing arrangement of subunits, connecting elements, key helices and key charged residues. Towards the end, the arrangement of peripheral domain is illustrated using T. thermophilus structure (PDB 3I9V) shown in grey with Fe-S clusters as spheres. (MOV 22129 kb)
Supplementary Movie 2
This movie illustrates the fold of antiporter-like subunits (using subunit NuoM), showing the fold from N- to C-terminus, key helices, key charged residues and internal symmetry. Two repeating domains are shown in magenta and green. (MOV 25622 kb)
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Efremov, R., Sazanov, L. Structure of the membrane domain of respiratory complex I. Nature 476, 414–420 (2011). https://doi.org/10.1038/nature10330
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DOI: https://doi.org/10.1038/nature10330
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