GPCR Solubilization and Stabilization
CD BioSciences provides professional services for GPCR solubilization and stabilization to facilitate the cutting-edge programs of GPCR research and drug development.
Introduction of GPCR Solubilization and Stabilization
GPCRs, a large family of membrane proteins with seven transmembrane domains, play a critical role in the regulation of many physiological processes and pathological conditions. A pre-requisite for the success of GPCR research and drug development is the ability to study the isolated receptor proteins that preserve the proper folding and pharmacology. However, the intrinsic instability in aqueous solutions and the hydrophobic nature of GPCRs make it difficult to isolate functional GPCRs from membrane without compromising the structural and functional integrity of the protein.
Detergents are widely used for GPCR solubilization, which is a key step to dissociate the membrane proteins and lipids in native membranes. After GPCR solubilization, it is often necessary to remove the detergents and reconstitute them to a defined stable lipid system to overcome the obstacles of the inherent instability of GPCRs in the detergents. A stabilized receptor, which is more conformationally homogeneous, will also facilitate crystallogenesis. Amphipathic materials, such as detergents, amphipols and nanodiscs, play an indispensable role in these steps.
Figure 1. A schematic representation of different stages of solubilization of biological membranes by detergents. (Chattopadhyay A, et al., 2015)
There is not one right amphipathic material for the solubilization and stabilization of each GPCR. The correct choice is crucial to maintain the functional state of solubilized receptors. In addition, the solubilization procedure must also be optimized to yield the highest extraction efficiency.
Equipped with powerful GPCR platforms and experienced scientists, CD BioSciences is willing to work closely with our customers to help find the most suitable strategies and contribute to GPCR projects with our unreserved services and best efforts.
|Detergents||Detergents are soluble amphiphiles with critical micelle concentrations (CMCs) typically in the range of millimolar. Detergents used for GPCR solubilization can be divided into ionic detergents, non-ionic detergents and zwitterionic detergents. A number of measures have been used to increase the stability of GPCRs in detergent solution.|
|Micelle||Micelles are amphiphilic colloidal structures formed by self-assembly of amphiphilic molecules. They can be used keep GPCRs homogeneously dispersed in aqueous solutions with intact intrinsic functions.|
|Liposomes||Liposomes with a lipid bilayer structure can be used to mimic cell membranes. Reconstitution of detergent-solubilized GPCRs into liposomes can help maintain the proper folding, native structures, and functions of GPCRs.|
|Amphipols||Amphipols, a class of amphiphilic polymers, can be used to preserve the native fold of GPCRs purified from membrane fractions and also maintain their pharmacological properties.|
|Nanodiscs||Nanodiscs, a synthetic model membrane system, is composed of membrane scaffold proteins and phospholipid molecules. GPCRs can be integrated into nanodiscs to maintain its biological activity.|
|Lipodisqs||Lipodisqs, a discoidal nanoscale lipid bilayer system, are formed by styrene maleic acid (SMA). SMA can be used to extract GPCRs directly from native membranes. It offers a detergent-free method for GPCR solubilization which retaining the annular lipid environment and maintaining the native structure of target protein.|
If you are interested in our services for GPCR solubilization and stabilization, please feel free to contact us for more details.
- Chattopadhyay A, et al. Solubilization of G protein-coupled receptors: a convenient strategy to explore lipid–receptor interaction. Methods in enzymology. Academic Press, 2015, 557: 117-134.
- Jamshad M, et al. G protein-coupled receptors solubilization and purification for biophysical analysis and functional studies, in the total absence of detergent. Bioscience reports, 2015, 35(2).
- Bertheleme N, et al. Unlocking the secrets of the gatekeeper: methods for stabilizing and crystallizing GPCRs. Biochimica et Biophysica Acta (BBA)-Biomembranes, 2013, 1828(11): 2583-2591.