Nanodiscs Construction
Membrane proteins (MPs) have long been a challenge to biochemical and functional studies because they often display altered or loss of activity and function outside of a phospholipid environment. Nanodiscs, by providing a native membrane environment, have proven to be an invaluable tool for revealing the structure and function of MPs as well as their complexes with other proteins and lipids. A nanodisc is a synthetic membrane system composed of a lipid bilayer of phospholipids with the hydrophobic edge stabilized by two amphipathic helical protein belts, termed membrane scaffold proteins (MSPs). The size of nanodiscs is determined by the length of the MSPs and the stoichiometry of the lipids used in the self-assembly process. Nanodiscs are useful in the study of membrane proteins because they can solubilize and stabilize membrane proteins and represent a more native environment than liposomes, detergent micelles, bicelles and amphipols.
Creative Biostructure has a professional team of scientists who have been working with membrane proteins for years, and are actively establishing and improving related techniques within the membrane protein screening platform. Using the nanodiscs technology, our scientists have successfully incorporated various membrane protein targets of different topologies into the nanodisc bilayer to preserve MP structure and activity. By searching for an optimal molar stoichiometric ratio of lipid to scaffold protein as well as scaffold protein to MP (usually detergent-solubilized), the composition and homogeneity of the resultant assembly can be optimized to maximize the yield of MP incorporation. Membrane proteins prepared in nanodiscs are compatible with X-ray crystallography, EM and NMR studies.
Figure 1. Incorporation of membrane protein in nanodisc
Features of Our Service
- Combination of nanodiscs and cell-free or cell based MP production
- Uniform size, robust stability of nanodiscs and incorporated MPs
- Detergent-free environment that maintains MP function and activity
- Controlled oligomerization of membrane proteins and lipid compositions
- Compatibility with most protein purification methods
- Compatibility with many biochemical and analytical techniques developed for soluble proteins
Creative Biostructure takes advantage of the most advanced nanodisc technology to reconstitute membrane proteins in their native state and conformation. This greatly facilitates the structural/functional studies to understand the molecular mechanisms of membrane proteins and their interaction with other proteins, nucleic acids and lipids. Please see the list of membrane proteins provided by Creative Biostructure.
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References
Denisov, I.G. and Sligar, S.G. (2016) "Nanodiscs for Structural and Functional Studies of Membrane Proteins." Nature Structural and Molecular Biology 23, 481-486.
