Abstract
Nanodiscs are disc-shaped self-assembled lipid bilayers encircled by membrane scaffolding proteins derived from Apolipoprotein A-1 (apo A-1). They constitute a versatile tool for studying membrane proteins since reconstitution into nanodiscs allows studies of the membrane proteins in detergent-free aqueous solutions in a lipid bilayer. Here, we apply the technique to the Na+,K+-ATPase (NKA) from pig kidney using Membrane Scaffolding Protein 1 D1 (MSP1D1). Contrary to other reports, the nanodiscs obtained by our protocol are built up of the native lipids originally present in the detergent solubilized sample together with the NKA.
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References
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Acknowledgements
This protocol has been developed in collaboration with Linda Reinhard, Department of Molecular Biology and Genetics, Aarhus University. The expert technical assistance of Ms. Birthe Bjerring Jensen, Department of Biomedicine, Aarhus University, is gratefully acknowledged. J.L.G. was supported by a PhD stipend from the Centre for Membrane Pumps in Cells and Disease—PUMPKIN, Danish National Research Foundation, T.B. was supported from the ERC advanced grant—BIOMEMOS awarded to P.N.
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Gregersen, J.L., Fedosova, N.U., Nissen, P., Boesen, T. (2016). Reconstitution of Na+,K+-ATPase in Nanodiscs. In: Bublitz, M. (eds) P-Type ATPases. Methods in Molecular Biology, vol 1377. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3179-8_36
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DOI: https://doi.org/10.1007/978-1-4939-3179-8_36
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3178-1
Online ISBN: 978-1-4939-3179-8
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