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Reconstitution of Membrane Proteins in Phospholipid Bilayer Nanodiscs
Open accessAbstract
Self-assembled phospholipid bilayer Nanodiscs have become an important and versatile tool among model membrane systems to functionally reconstitute membrane proteins. Nanodiscs consist of lipid domains encased within an engineered derivative of apolipoprotein A-1 scaffold proteins, which can be tailored to yield homogeneous preparations of disks with different diameters, and with epitope tags for exploitation in various purification strategies. A critical aspect of the self-assembly of target membranes into Nanodiscs lies in the optimization of the lipid:protein ratio. Here we describe strategies for performing this optimization and provide examples for reconstituting bacteriorhodpsin as a trimer, rhodopsin, and functionally active P-glycoprotein. Together these demonstrate the versatility of Nanodisc technology for preparing monodisperse samples of membrane proteins of wide-ranging structure.
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Cites
Structure of P-Glycoprotein Reveals a Molecular Basis for Poly-Specific Drug Binding
Aller, S. G., Yu, J., Ward, A., Weng, Y., Chittaboina, S., Zhuo, R., Harrell, P. M., Trinh, Y. T., Zhang, Q., Urbatsch, I. L., et al. (2009) · Science · doi:10.1126/science.1168750 · Cited by 1,929
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Cryo-EM structures of a human ABCG2 mutant trapped in ATP-bound and substrate-bound states
Manolaridis, I., Jackson, S. M., Taylor, N. M. I., Kowal, J., Stahlberg, H., & Locher, K. P. (2018) · Nature · doi:10.1038/s41586-018-0680-3 · Cited by 228
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Qian, H., Wu, X., Du, X., Yao, X., Zhao, X., Lee, J., Yang, H., & Yan, N. (2020) · Cell · doi:10.1016/j.cell.2020.05.020 · Cited by 185
Structural and mechanistic basis of the EMC-dependent biogenesis of distinct transmembrane clients
Miller-Vedam, L. E., Bräuning, B., Popova, K. D., Schirle Oakdale, N. T., Bonnar, J. L., Prabu, J. R., Boydston, E. A., Sevillano, N., Shurtleff, M. J., Stroud, R. M., et al. (2020) · eLife · doi:10.7554/elife.62611 · Cited by 87
A YidC-like protein in the archaeal plasma membrane.
Borowska, M. T., Dominik, P. K., Anghel, S. A., Kossiakoff, A. A., & Keenan, R. J. (2015) · Structure · doi:10.1016/j.str.2015.06.025 · Cited by 71
Metadata
- Authors
- Ritchie, T. K., Grinkova, Y. V., Bayburt, T. H., Denisov, I. G., Zolnerciks, J. K., Atkins, W. M., & Sligar, S. G.
- Venue
- Methods in Enzymology
- Publisher
- Elsevier
- Published
- 2009
- Volume
- 464
- Pages
- 211-231
- ISSN
- 0076-6879 (ISSN-L)
- OpenAlex
- W1598629441
- Cited by
- 872
- References
- 1
- FWCI
- 21.65
- Type
- book chapter
- Language
- en
- Open access
- green
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- Ritchie, T. K., Grinkova, Y. V., Bayburt, T. H., Denisov, I. G., Zolnerciks, J. K., Atkins, W. M., & Sligar, S. G. (2009). Reconstitution of Membrane Proteins in Phospholipid Bilayer Nanodiscs. Methods in Enzymology. https://doi.org/10.1016/s0076-6879(09)64011-8
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