Nanodisc MSP1D1 dH5-His DMPG Biotinyl PE (50 µl)

Order number: 26421

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Phospholipids

DMPC

POPC

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Description

Membrane scaffold proteins derive from apolipoprotein (apo; A-I). They are used in membrane protein sciences for the stabilization of membrane proteins in a lipid environment after the removal of the cell membrane. The size of the nanodisc is determined by the MSP that was used to create the disc. MSP1D1 dH5 Biotinyl PE nanodiscs have a ~7-8 nm diameter.

This product is a pre-assembled nanodisc. Its intended purpose is to stabilize a cell-free expressed membrane protein. There are multiple phospholipids to choose from that can make up the body of the nanodisc. Which is best, depends on your protein.

For the assembly of this nanodisc product, a mixture of 10% headgroup-biotinylated Phospholipid in DMPC or POPC has been used. The resulting nanodisc-protein complexes can be used e.g. for immobilization on streptavidin-coated surfaces.

Other MSP products or related sites by Cube Biotech include:

MSP nanodisc schematic overview — **Figure 1:** Schematic of a MSP nanodisc. A Membrane-scaffold-protein (MSP, green) forms a disc-shaped structure with some phospholipids.

Datasheets

Feature (includes data about the MSP protein)
Purity	> 90% (Determined by SDS-PAGE)
Number of amino acids	184
Molecular Mass	21,468 Da
Extinction coefficient (in water) ε₂₈₀	19,940 M^-1cm^-1
Buffer	20 mM TRIS pH 7.4; 100 mM NaCl; 0.5 mM EDTA
Origin	Human
Sequence (1-Letter Code), for more details, see datasheet	MGSSHHHHHH ENLYFQ GSTFSKLREQ LGPVTQEFWD NLEKETEGLR QEMSKDLEEV AKVQPYLDDF QKKWQEEMEL RQKVEPLGEE MRDRARAHVD ALRTHLAPYS DELRQRLAAR LEALKENGGA RLAEYHAKAT EHLSTLSEKA KPALEDLRQG LLPVLESFKV SFLSALEEYT KKLNTQ
Affinity tag	N-Terminal His-tag
Shipping Temperature	Dry-Ice
Storage of reconstituted protein	-80°C, avoid freeze-thaw cycles
Helpful literature references	Roos, C., Kai, L., Haberstock, S., Proverbio, D., Ghostastider, U., Ma, Y., Filipek, S., Wang, X., Dötsch, V., and Bernhard, F. (2014) High level cell-free production of membrane proteins with nanodiscs. Meth. Mol. Biol. 1118, 109-30 Roos, C., Zocher, M., Müller, D., Münch, D., Schneider, T., Sahl, H.G., Scholz, F., Wachtveitl, J., Ma, Y,. Proverbio, D., Henrich, E., Dötsch, V. and Bernhard, F. (2012) Characterization of co-translationally formed nanodisc complexes with small multidrug transporters, proteorhodopsin and with the E.coli MraY translocase. Biochim. Biophys. Acta 1818, 3098-106.

Lab Results

PRE-Assembled MSP nanodisc in use — **Figure 1:** Stability and specific activity of E. coli MraY translocase in different lipid environments. The protein was cell-free synthesized in E. coli lysates in presence of pre-assembled MSP1E3D1 nanodiscs containing the indicated lipids. The topology of the membrane-integrated MraY enzyme is indicated in the inset. MraY catalyzes the lipid-I precursor formation of bacterial cell wall biosynthesis. The figure shows the lipid-I formation in nmol per µg catalyzed by the corresponding MraY/nanodisc samples (Roos et al., (2012) BBA 1818, 3098-3106). The activity of the enzyme strongly depends on anionic phosphatidylglycerol type lipids and was best with the shorter myristoyl fatty acid chains (DMPG). Data kindly provided by Frank Bernhard, University Frankfurt.

PRE-Assembled MSP nanodisc in use II — **Figure 2:** Differential ligand binding activity of the human endothelin A (ETA) and endothelin B (ETB) receptors in MSP1E3D1 (DMPC) nanodiscs. The G protein-coupled receptors (GPCRs) ETA and ETB were cell-free synthesized in E. coli lysates in presence of pre-assembled MSP1E3D1 nanodiscs containing the lipid 1,2-dimyristoyl-sn-glycerol-3-phosphocholine (DMPC). The resulting receptor/nanodisc samples were analyzed for their characteristic binding affinities to four different peptide ligands by SPR (Proverbio et al., (2013) BBA 1828, 2182-2192). Both GPCRs have high affinity to the natural ligand ET-1 while the affinities to the other three ligands are receptor specific. The characteristic differential ligand binding profile of the cell-free synthesized GPCR/nanodisc samples was obtained and in good agreement with data from the literature. Data kindly provided by Frank Bernhard, University Frankfurt.

Video

Watch our video tutorial about the creation of MSP nanodiscs. It explains the whole process, starting with the initial solubilzation using detergents. The protein that was stabilized in this demonstration was Bacteriorhodopsin.

We also recommend our video guide to nanodiscs in general.

FAQ

Can I get the datasheet for the pre-assembled MSP1D1 dh5 (His-tagged) Biotinyl PE nanodiscs?

Yes, just download them here.

Is this the correct MSP product for my membrane protein?

This cannot be answered easily as every membrane protein performs differently with each nanodisc size and phospholipid. It usually requires a screening process before. Remember that membrane proteins with fewer transmembrane domains (TMD) require a smaller nanodisc, while a high number of TMDs need a larger nanodisc.

What the correct phospholipid for your membrane proteins is, is also protein dependent, unfortunately.

I have never worked with MSP nanodiscs before. Can you tell me more about them?

Of course! In fact we created THIS MSP NANODISC GUIDE PAGE, for this purpose. Have a look at it!

Are there other options to stabilize membrane proteins besides MSP nanodiscs?

Yes, there are. We recommend synthetic copolmer nanodiscs greatly! Have a look at THIS GUIDE PAGE to learn more.

Disclaimer

Our products are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.
Nanodiscs are protected by US Patents 7,691,414; 7,662,410; 7,622,437; 7,592,008; 7,575,763; 7,083,958; 7,048,949