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Based on the advanced Mempro™ Nanodisc preparation platform established for a long time, Creative Biostructure can offer high-quality Mempro™ Nanodisc design services to meet all your demands.
Nanodiscs are soluble nanoscale phospholipid bilayers, which are encircled and stabilized by membrane scaffold proteins (MSP). They have been proved to be a powerful and promising tool to study membrane proteins. The MSP used for generating Nanodiscs is based on the sequence of human serum apolipoprotein AI. A bilayer domain is formed by the combined phospholipid, meanwhile, the edges of the discoidal structure are warped around by two molecules of MSP in a belt-like configuration, one MSP covering the hydrophobic alkyl chains of each leaflet. Creative Biostructure has focus in Nanodisc design services for years, our services are full-scale including the nanodisc size, choice of phospholipid&detergent, determination of phospholipid/MSP molar ratio, etc.
Figure 1. Formation of Nanodiscs occurs by mixing membrane scaffold protein with detergent solubilized phospholipids. (Lab on a Chip 2008)
Generally, the size of an empty nanodisc can be modulated from 9.5 nm to 17 nm. Two essential factors should be taken into account: the size of the transmembrane portion of the target protein that needs to be reconstituted in the nanodisc, and the number of target membrane proteins required to be combined.
It’s the native cell type that determines the lipid ingredient surrounding of membrane proteins. To choose phospholipid for nanodisc reconstitution, there are three factors need to be considered: the molecular shape, the lipid composition and the phase transition temperature (Tm).
The detergent used for initial solubilization is important as well, usually is sodium cholate which is perfect for the lipid fraction and is generally present at a 2:1 mole ratio to total PL, or higher.
In order to get high return homogeneous Nanodiscs, a specific ratio of phospholipid to MSP is necessary during the assembly process based on the MSP belt-length/Nanodisc diameter. The presence of the membrane protein needs a decrease in the number of phospholipid molecules, in contrast to an empty-nanodisc, according to the size of the transmembrane region of the membrane protein.
Inagaki S, Ghirlando R, Grisshammer R. Biophysical characterization of membrane proteins in nanodiscs[J]. Methods, 2013, 59(3): 287-300.
Shih A Y, Freddolino P L, Arkhipov A, et al. Assembly of lipoprotein particles revealed by coarse-grained molecular dynamics simulations[J]. Journal of structural biology, 2007, 157(3): 579-592.
Denisov I G, Grinkova Y V, Lazarides A A, et al. Directed self-assembly of monodisperse phospholipid bilayer Nanodiscs with controlled size[J]. Journal of the American Chemical Society, 2004, 126(11): 3477-3487.
Yang J, Liu H, Martens W N, et al. Synthesis and characterization of cobalt hydroxide, cobalt oxyhydroxide, and cobalt oxide nanodiscs[J]. The Journal of Physical Chemistry C, 2009, 114(1): 111-119.