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Membrane Protein Purification Using Nanodiscs

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Creative Biolabs provides advanced nanodiscs-based membrane protein purification service. We are capable of satisfying all your specific demands through the whole project.

Nanodiscs, the nanoscale systems (the average size from 9.5 to 12.8 nm) with self-assembling ability, can stabilize membrane proteins extracted from the membrane with surfactants. Nanodiscs can also provide a phospholipid bilayer system maitained by the amphipathic membrane scaffold proteins (MSPs) (Figure 1), which are truncated forms of human apolipoprotein A-I (apo A-I) that wrap around the hydrophobic core of the lipids to form a disc-like particle.

Figure 1. Model of P-glycoprotein incorporated into 9.8-nm nanodisks. Blue: Membrane scaffold proteins (MSPs), Grey: Phospholipids, Pink: Reconstituted P-glycoprotein.

Nanodiscs have sufficient and unique advantages compared with other membrane protein purification approaches: It can provide an excellent and nearly natural environment to keep membrane proteins stable. Thus, nanodiscs can be applied in a wide range of utilities, including:


In addition, nanodiscs can also be used to reconstitute membrane proteins such as GPCRs. Briefly, the nanodisc-solubilized proteins can be directly purified without the use of detergents. The completed assemblage allow for the access to the physiologically intracellular and extracellular faces of the protein, which is its unique characteristic suit to G proteins and other types of protein with their own interaction partners.

Up to now, there are two methods for reconstituting membrane proteins into nanodiscs:
1). Solubilizing and purifying the membrane proteins with a suitable detergent, followed by the addition of membrane scaffold proteins and phospholipids into the mixture. Subsequentially, the nanodiscs containing the membrane proteins will assemble automatically. For more information, please click here .
2). Expressing the membrane proteins in a cell-free system with the addition of pre-assembled nanodiscs that contains the integrated membrane proteins. For more information, please click here.

In the latter method, not only the detergent is not needed, moreover, the cell-free expression system also provides the possibility to introduce modifications into membrane proteins, such as biotinylation or isotope labelling. According to your specific project, we can evaluate and select either of these two options to reconstitute membrane proteins into nanodiscs.

Creative Biolabs can provide all types of membrane scaffold proteins and phospholipids to reconstitute your own membrane proteins into nanodiscs. We also provide other membrane protein production services. Please feel free to contact us for a detailed quote.

References:
A. Leitz et al. (2006). Functional reconstitution of β2-adrenergic receptors utilizing self-assembling Nanodisc technology. BioTechniques. 40(5): 601-606.
A. Robinson et al. Production of Membrane Proteins.
R. M. Bill, et al. (2011). Surfactant-free purification of membrane proteins with intact native membrane environment. Biochem. Soc. Trans. 39: 813-818.




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