Membrane Protein Services
Our represented companies offer services related to transmembrane proteins, from small to medium scale, for a wide range of applications. The service is modular, it consists of 6 modules and you can choose a complete service, as well as one or more of the services.
This service is useful for: ligand detection and diagnostic assays, cryoelectron microscopy, antibody generation, and crystallization, among others.
Module 1 – Protein Expression
In addition to expression by E.coli, they are capable of using three different expression systems:
- Cell free expression system: : A perfect match with pre-assembled nanodisc for membrane protein expression since it is an open system with no cell membrane. The membrane protein is co-translationally integrated into nanodisc. Recommended for toxic proteins and membrane proteins that need to be stabilized in all types of nanodiscs.
- Baculovirus Insect system: Near mammalian like post-translational modifications. Easy to handle eucaryotic expression system.
- Hek293 cells : For homologue human protein expression and comprehensive post-translational modifications. We offer the BacMam system and transient transfection in Hek293 suspension cultures.
Module 2 – Protein solubilization
This step is essential to obtain strucutral intact and functional membrane proteins. Two different methods can be chosen for this step:
- Ultrapure detergents: Detergents are the classic approach to solubilize membrane proteins.
- Synthetic Polyers: Polymers like SMA and DIBMA can form complexes that are called synthetic nanodiscs. Their key advantage: They combine the essential steps of membrane proteins solubilization and subsequent stabilization. This fuses our modules 2 and 3.
Module 3 – Protein Stabilization
The goal is to stabilize membrane proteins and keeping them functional after their native cell membrane has been removed. The main idea is to mimic the original cell membrane of the membrane protein of interest to ensure maximal authenticity with its functions. Three different options:
- Detergents: The traditional way to stabilize membrane proteins. We offer a great choice of different detergents.
- MSP Nanodiscs: With their controlled phospholipid composition, a near perfect replication of the native membrane enviroment of the membrane protein can be created.
- Synthetic Nanodiscs: As an innovative way, the synthetic polymer DIBMA has the capability to solubilize and stabilize a membrane protein. Using Synthetic polymers like DIBMA leave the original membrane enviroment of the membrane protein of interest intact.
Module 4 – Protein Purification
To extract the desired protein from the rest of the cell’s components after cell lysis, there are several options too:
- Surface Affinities/Affinity tags
- Customized agarose resins / magnetic beads
Module 5 – Protein Characterization
Now it is time to identify the characteristics of your protein of interest. For this we offer:
- Dynamic light scattering (DLS)
- Surface Plasmon resonance (SPR)
- Protein Stability Assays
Module 6 – Protein Structure Determination
One of the key characteristics of a protein is its 3D structure. We offer two methods for it, Cryo-EM and Cubic Phase Crystallization.
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