Extraction of Membrane protein form EAEC requires to remove all the lipid contents. What should I can use for this purpose.
Are you trying to solubilize a single, recombinantly expressed membrane protein or total membrane proteins from E. coli?
Solubilizing over-expressed membrane proteins for purification should be empirically tested using different detergents, possibly their mixtures and different time points. This way you should be able to determine optimal solubilization conditions for you protein, where the protein is quantitatively separated from membranes in a folded and biologically active form. Triton X works sometimes, but I have usually just used it more like a control, similarly to SDS, when trying out more elegant detergents like DM, DDM, Fos-Choline 12 or similar.
Triton X and SDS can be used to disrupt membranes for crude isolation of total membrane proteins, although these detergents may also cause proteins to denature.
Hi, triton X-100 is used to solubilize membrane proteins but it by itself do not remove lipids. They will probably remain in your extract forming micelles. I think that the best choice to remove lipids is precipitating proteins with any solvent like chloroform or acetone.
It depends on your downstream application. TX100 is a quite harsh detergent, such as SDS. You might also consider using milder detergents, such as CHAPS or polyoxyethylenes (POE), which could preserve the structure of several membrane proteins.
However, you would be solubilizing the proteins by substituting the lipids. If you want complete removal of lipis and do not care about protein structure and/or interactions, you can simply precipitate the proteins with TCA, for example.
Triton X can aggregate the proteins that you are trying to isolate.Studying membrane proteins represents a major challenge in protein biochemistry, with one of the major difficulties being the problems encountered when working outside the natural lipid environment. In vitro studies such as crystallization are reliant on the successful
solubilization or reconstitution of membrane proteins, which generally involves the careful selection of solubilizing detergents and mixed
lipid/detergent systems. So, focus on the relevant
molecular properties of the detergents and lipids that aid understanding of these processes. A significant barrier to membrane protein research
is retaining the stability and function of the protein during solubilization, reconstitution and crystallization.
You can try out, DDM, n-dodecyl-h-d-maltoside; DDAO, dodecyldimethyl-N-amineoxide; DMPC, 1,2-dimyristoyl-sn-glycero-3-phosphocholine;
CHAPS, 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate; MGDG, monogalactosyl diacylglycerol; DGDG, digalactosyl diacylglycerol;
DPPG, l-a-dipalmitoylphosphatidylcholine; DOPG, l-a-dioleoylphosphatidylglycerol; DOPC, l-a-1,2-dioleylphospatidylcholine; DOPE, l-a-1,2-dioleylphospatidylethanolamine; DHPC, l-a-1,2-dihexanoylphosphatidylcholine; DAG, diacylglycerol; POPC, 1-palmitoyl-2-oleoyl-phosphatidylcholine;
CHAPSO, 3-[(3-cholamidopropyl)dimethylammonio]-2-hydroxyl-1-propanesulfonate
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Proteins