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Magic™ Membrane Protein Incorporation in Nanodiscs

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Creative Biolabs now provides the professional custom service in membrane protein incorporation in nanodiscs. Lipid-bilayer nanodiscs are the most powerful tool in solubilizing/incorporating membrane proteins in a cell membrane-like environment. This solubilization/incorporation system represents a very suitable option if you need to keep the folded conformation, intact function and oligomerization of your target proteins all in their native forms.

Usually, membrane proteins can be solubilized, reconstituted or incorporated in amphipols, micelles, bicelles, liposomes, and nanodiscs. Compared to the other systems, nanodiscs can offer a series of advantages including:


However, incorporating membrane proteins into nanodiscs is not an easy task to accomplish. Our scientists have comprehensive knowledge and years of experience in handling membrane proteins with nanodisc lipid bilayer particles. We guarantee that the membrane protein-incorporated nanodiscs prepared by us will be


Assembled Nanodiscs

Lipids:
For preparing assembled nanodiscs, zwitterionic lipids or a mixture of zwitterionic and negatively charged lipids are commonly used. At Creative Biolabs, a large variety of such lipids are available on our lipid screening platform. Our scientists can always find the one that is most suitable for your membrane proteins based on our pre-analysis on the feature and native environment of your membrane proteins. Upon request, we can also perform a screening test of lipids, lipid mixtures, and soluble and lipid-bilayer additives to make sure that the stability and native function of your membrane proteins will be maximally maintained.

 

MSPs:
Another key component for preparing assembled nanodiscs is the membrane scaffolding protein (MSP). MSP is a belt-like protein winding around and stabilizing the nanodisc. We provide apolipoprotein A-1 based MSPs at different lengths to match the specific needs of our customers. By adjusting the length of the MSPs, we guarantee that the diameter of the nanodiscs prepared by us will match the requirements of our customers in order to fix the membrane protein/lipid ratio and "lock" the membrane proteins in a specific oligomerization state.

Magic™ Membrane Protein Incorporation in NanodiscsFigure 1. Model of P-glycoprotein incorporated into 9.8-nm nanodiscs.
Blue: membrane scaffold proteins (MSPs), Grey: phospholipids, Pink: reconstituted P-glycoprotein.

In Creative Biolabs, assembled nanodiscs can be prepared in either the detergent-assistant or detergent-free method.

In the detergent-assistant method, we can start with the membrane proteins provided by you or synthesized by us via either cell-based protein expression or cell-free protein expression. After that, we will first solubilize your membrane proteins using detergents and purify them in a micellar solution. Since different detergents and detergent mixtures may affect the stability, structure, function, and activity of a membrane protein very differently, we can also use our detergent screening platform to find the detergent or detergent mixture most suitable for your membrane proteins upon your request. After the solubilization and purification, we will add lipids and the membrane scaffolding protein (MSP) to the micellar solution of your membrane proteins followed by removing the detergents via hydrophobic beads binding and dialysis. At the same time, the assembled nanodiscs with your membrane proteins embedded will form.

In the detergent-free method, we will combine the nanodisc incorporation service with our cell-free protein expression technique to make sure your membrane proteins will be directly expressed and folded into the assembled nanodiscs in a cell-free expression system. For more information, please consult our service in cell-free membrane protein preparation in nanodiscs. Since detergent micelles may not be a good mimic of cell membranes, solubilization of some membrane proteins in detergent micelles may cause their unfolding, conformational change, loss of function and/or deactivation. In that case, use of detergents shall be avoided during the whole procedure of the membrane protein-lipid nanodisc preparation.

"Native" Nanodiscs

In Creative Biolabs, "native" nanodiscs can also be prepared by extracting your membrane proteins together with their surrounding lipids and other components in the cell membrane after cell-based expression with styrene-maleic acid copolymers (SMA). Like the MSP, SMA can work as the "belt" winding around and stabilizing the nanodiscs. In this method, we will not only prepare your nanodiscs without using any detergents but also avoid adding any exogenous lipids. Your nanodiscs will be composed of all native components in the cell membrane except the SMA, and therefore the native conformation, stability, function, and activity of your membrane proteins will be almost 100% maintained in these "native" nanodiscs.

Finally, we will purify the nanodiscs using tag-based and/or gel-filtration chromatography. Affinity tags can be added to your membrane proteins and/or the MSP. Besides, you can also choose our lyophilizing or freezing service as an add-on option for the long-term storage of the nanodiscs.

Magic™ Membrane Protein Incorporation in Nanodiscs

Nanodiscs are almost the perfect tool for solubilizing, reconstituting and incorporating membrane proteins in a cell membrane-like environment. At Creative Biolabs, we guarantee our clients the most qualified and tailored service to satisfy each specific demand. Besides, you can also choose our services in solubilization/reconstitution of membrane proteins in liposomes, micelles, and amphipols. Please feel free to inquire us for more information.




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