(like SDS, n-octyl-β-d-glucopyranoside (OG)
, n-dodecyl-β-d-maltoside (DDM)
are widely used in membrane protein solubilization even though it is well known that different detergents have different weaknesses. Short chain nonionic detergents for example can affect the functional properties of a membrane protein (1). It seems clear that removing the native lipid bilayer from the membrane protein can interfere with the function of the protein. One way to mimic the native lipid membrane are MSP-nanodiscs
(Fig. 1) or detergent-free polymer systems (Fig. 2) (Styrene-maleic acid co-polymers (SMAs) (2), Diisobutylene-maleic acid (DIBMA)) (3,4). With the latter you can directly extract membrane proteins from cells without an intermediate step of detergent solubilization (5). Synthetic polymers have to carry a styrene or maleic acid group himself to solubilize proteins.
Fig.1: Nanodisc with a target protein and membrane scaffold proteins (MSPs, green).
Fig.2: Synthetic Polymers like SMA or DIBMA carrying a target protein.
Why it is advantageous to use PureCube DIBMA and not SMAs for your protein solubilization?
SMAs have big advantages in contrast to many detergents and are succsessfully used for many applications (6). The drawback of SMAs is a high absorbance of ultraviolet light in solution with an absorbance maxima at 280 nm. The main reason for this peak at 280 mn are aromatic amino acids like tryptophan or phenylalanine, SMA itself carries an aromatic ring. So quantitative protein concentration measuring of your sample is not possible during the process of membrane protein solubilization, stabilization and purification. With PureCube DIBMA from Cube Biotech you can solubilize, stabilize and purify your protein detergent-free and measure your protein concentration without any problems.
Fig.3: SDS Page showing solubilized membrane-protein fractions of E. coli. 10 mM (0.5 % (w/v)) DDM and 3 mM (2.5 % (w/v)) DIBMA was used. The figure is taken from Grethen et. al. 2017 (7).
Fig.4: Chemical structural formula of diisobutylene-maleic acid (DIBMA).
Why you should use PureCube DIBMA from Cube Biotech?
We provide our product highly purified and lyophilized. On top of that, PureCube DIBMA is lyophilized from two different buffer solutions (HEPES or TRIS) to ensure a stable pH at 7.5 which is ideal for most protein solubilizations. Feel free to contact
us if you wish to have PureCube DIBMA in a different kind of buffer composition. For different applications you can choose from samples with a medium length of 10,000 or 12,000 g/mol.
MW: ~12,000 g/mol or 10,000 g/mol
dn/dc: 1.35 M-1
Solubility: > 10 % in H2O
Absorbance at 280 nm (1 % solution): < 0.3
Mg2+ Tolerance: 25 mM
Ca2+ Tolerance: 20 mM
1. Seddon, Annela M., Paul Curnow, and Paula J. Booth. "Membrane proteins, lipids and detergents: not just a soap opera." Biochimica et Biophysica Acta (BBA)-Biomembranes 1666.1-2 (2004): 105-117.
2. Knowles, Timothy J., et al. "Membrane proteins solubilized intact in lipid containing nanoparticles bounded by styrene maleic acid copolymer." Journal of the American Chemical Society 131.22 (2009): 7484-7485.
3. Oluwole, Abraham Olusegun, et al. "Solubilization of Membrane Proteins into Functional Lipid‐Bilayer Nanodiscs Using a Diisobutylene/Maleic Acid Copolymer." Angewandte Chemie International Edition 56.7 (2017): 1919-1924.
4. Oluwole, Abraham Olusegun, et al. "Formation of lipid-bilayer nanodiscs by diisobutylene/maleic acid (DIBMA) copolymer." 33.50 (2017): 14378-14388.
5. Long AR, O’Brien CC, Malhotra K, Schwall CT, Albert AD, Watts A, Alder NN (2013) A detergent-free strategy for the reconstitution of active enzyme complexes from native biological membranes into nanoscale discs. BMC Biotechnol 13:41
6. Dörr, Jonas M., et al. "The styrene–maleic acid copolymer: a versatile tool in membrane research." European Biophysics Journal 45.1 (2016): 3-21.
7. Grethen, Anne, et al. "Thermodynamics of nanodisc formation mediated by styrene/maleic acid (2: 1) copolymer." Scientific reports 7.1 (2017): 11517.