Mempro™ Nanodisc Assembly Kits
Creative Biostructure has committed to the development of the novel nanodisc technology in recent years, and has developed a magic platform named Mempro™ Nanodisc to provide all kinds of products and services. Mempro™ Nanodisc Assembly Kits can help you to generate custom empty nanodiscs where nascent membrane proteins can be inserted directly without detergent extraction.
Figure 1. Illustration of the self-assembly process of nanodiscs (Mater Methods, 2013).
The nanodisc self-assembly is a so-called directed self-assembly process. It is a two-step process where the phospholipids are first dissolved by a detergent into mixed lipid-detergent micelles and then the mixed micelles are transformed into phospholipid bilayer discs by incubating with MSP and subsequently removing the detergent using biobeads. In this context the amphiphatic MSP may also be considered a kind of detergent that will distribute itself between the solution and lipid-detergent micelles. During the incubation the mixed lipid-detergent micelles will bind some MSPs, which will be incorporated into the lipid-detergent micelle. The added Biobeads are small porous polymer beads that will trap detergents, effectively removing them from solution and in turn removing them from the mixed micelles. The removal of the detergents changes the effective packing parameter of the detergent-lipid micelles from spherical to a planar bilayer geometry, which in turn is stabilized by the MSPs. When the mixing stochiometries are right, the majority of the particles formed will be nanodiscs containing 130-150 phospholipids. The formed nanodiscs are finally separated form larger aggregates and excess MSPs using size exclusion chromatography.
Creative Biostructure can provide both the aliquots and protocols of Mempro™ Nanodisc Assembly Kits for your convenience. We recommend you to use these Mempro™ Nanodisc Assembly Kits with membrane proteins expressed by the cell-free system because they can be directly inserted into nanodiscs without the prior process of detergent extraction. In our Mempro™ Nanodisc Assembly Kits, the MSP/lipid ratio is the optimized 1:80. Our Mempro™ Nanodisc Assembly Kits are aliquots of MSPs, phospholipids, and sodium cholate which are ready to use and very ideal for combination of cell-free expression reactions. Mempro™ Nanodisc platform will also provides the expert support and protocols in details.
Creative Biostructure can provide a series of Mempro™ Nanodisc Assembly Kits composed by different membrane scaffold proteins and different phospholipids. You can use these kits to achieve custom nanodiscs of varying sizes with the desired membrane proteins. Creative Biostructure also provide His-tagged MSPs to allow the easy detection.
Mempro™ Nanodisc Technology is a magic tool to investigate the structure and function of membrane proteins, and there are a wide range of applications based on it. Creative Biostructure can also offer other Mempro™ Nanodisc products. Please see the related sections below for more information and feel free to contact us for a detailed quote.
References:
P. K. Dominik, et al. (2016). Conformational Chaperones for Structural Studies of Membrane Proteins Using Antibody Phage Display with Nanodiscs. Structure,24: 300–309.
I. G. Denisov and S. G. Sligar. (2016). Nanodiscs for structural and functional studies of membrane proteins. Nature Structural & Molecular Biology,23: 481–486.
N. Mitra. (2013). Nanodiscs: Membrane Protein Research in Near-Native Conditions. Mater Methods, 3:177
A. Y. Shih, et al. (2007). Disassembly of nanodiscs with cholate. Nano Letters, 7(6):1692–1696.