Creative Biostructure

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Liposome Construction

Liposomes are artificial spherical vesicles with enclosed phospholipid bilayer structures. They are most often composed of phospholipids, especially phosphatidylcholine, but may also include other lipids such as egg phosphatidylethanolamine, as well as pure surfactant components like DOPE (dioleoylphosphatidylethanolamine). Liposomes are now an established technology platform with considerable clinical acceptance for the delivery of anti-cancer, anti-fungal, anti-inflammatory and antibiotic drugs, as well as nucleic acid-based therapeutics. Besides, liposomes are also frequently used as supplements to assist membrane protein synthesis, forming proteoliposomes, which serve as a model for the study of many membrane proteins.

Creative Biostructure has established proteoliposomes as a powerful tool to study the function of membrane proteins. With the membrane protein screening platform, our scientists at Creative Biostructure have been optimizing different techniques to insert proteins into liposomes, including mechanical means, freeze-thawing, organic solvents, and detergent-mediated method. The detergent-mediated pathway is most frequently used at Creative Biostructure due to its simplicity and robustness.

Solvation of membrane protein  in liposome
Figure 1. Solvation of membrane protein in liposome

Features of our service

  • Protein expression: Either cell-free translation system or cell-based system with the aid of solubilizing detergent is used for membrane protein production.
  • Mixture preparation: An excess of detergent-destabilized liposomes is added to form a solution of mixed phospholipid–protein–detergent complexes.
  • Proteoliposome reconstitution: Detergent is removed from micellar solutions using various strategies including dialysis, gel filtration, dilution and polystyrene beads (such as Amberlite XAD and Bio-Beads SM-2).
  • Efficiency analysis: Reconsititution efficiency should be evaluated by measuring the activity of the proteins, or by other methods available at Creative Biostructure.

Creative Biostructure takes advantage of the most advanced liposome technology to reconstitute membrane proteins in their native state and conformation. This greatly facilitates the structural/functional studies of transmembrane or membrane-associated proteins. Please see the list of membrane proteins provided by Creative Biostructure.

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Ordering Process

Ordering Process

References

Claxton DP, et al. (2015) Navigating Membrane Protein Structure, Dynamics, and Energy Landscapes Using Spin Labeling and EPR Spectroscopy. Methods Enzymol 564:349-387.


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