Cryo-EM for Membrane Proteins
Membrane protein refers to the general term for proteins that can bind or integrate into the membrane of cells or organelles. As the gateway between cells and the surrounding environment, and cell-to-cell material, information, energy exchange, membrane proteins play an essential role in physiological and biochemical activities, such as signal transduction, transmembrane transport of substances, cell-to-cell communication, and migration. Membrane proteins are the main targets for the treatment of human diseases. Currently, 60% of drug targets are membrane proteins, and the market share of drugs targeting membrane proteins is also increasing yearly. However, the structural analysis of membrane proteins has always had a bottleneck that is difficult to break through. According to the statistics of Protein Data Bank, membrane protein structures account for only 3% of the successfully analyzed three-dimensional protein structures, which is a huge difference from the fact that membrane protein genes account for 1/3 of the total number of genes in organisms. There are huge challenges in the current structural research of membrane proteins.
Our Cryo-EM Technology for Membrane Proteins
Cryo-electron microscopy (cryo-EM) is a powerful and rapidly developing structural biology technique that enables the visualization of biomolecules and complexes at near-atomic resolution. Cryo-EM has made rapid progress in analyzing the structure of complex membrane proteins, and a large number of high-resolution structures have been successfully resolved. It is an attractive method for studying the structural biology of membrane proteins. Creative Biostructure developed an optimized cryo-EM platform and established a method to fuse GPCR soluble regions with structures to efficiently determine the structures of GPCRs and their complexes with ligands/drugs.
Figure 1. Cryo-EM structures of different G protein-coupled receptors.
Our Service Process for Obtaining Membrane Protein Cryo-EM Structures
| procedure | Information |
| 1 | Construction of expression vector |
| 2 | Cell overexpression and protein purification |
| 3 | Complex formation with small molecules, frozen sample preparation |
| 4 | Preliminary collection and processing of cryo-electron microscopy data, screening and judging the quality of samples |
| 5 | High-quality cryo-EM data collection |
| 6 | Data processing to reconstruct high-resolution electron densities |
| 7 | Building and refining protein structure models to obtain high-precision complex structures |
Advantages of Our Membrane Protein Cryo-EM Services
Creative Biostructure has rich experience in membrane protein research, and the advantages of our service include:
- One-stop service: we provide one-stop membrane protein cryo-EM services, including plasmid design, protein expression, solubilization, purification, reconstruction, and structure determination.
- Variety of sample types: we have extensive experience in processing a variety of protein samples, including GPCRs, ion channels, transporters, receptor tyrosine kinases, and biologics (Fab-antigen structures).
- Advanced equipment: we have the most advanced electron microscope, optimized cryo-electron microscope platform, and powerful data processing software.
- Optimizing experimental methods: we optimize the results of cryo-electron microscopy through various methods, such as using detergent to expand the membrane protein and rapid visualization of large membrane proteins using negative staining TEM.
- Single-particle reconstruction to build 3D structural models of membrane proteins at high resolution.
Creative Biostructure promises to work closely with our customers to provide tailored EM strategies for the membrane proteins of interest. The result of this service, when combined with results from other biophysical and biochemical characterization methods, can provide detailed information that will ultimately facilitate the drug design process and the engineering of membrane proteins with novel functions for biotechnological applications. Please feel free to contact us for a detailed quote.
Ordering Process
Reference
- Piper S J., et al. Membranes under the Magnetic Lens: A Dive into the Diverse World of Membrane Protein Structures Using Cryo-EM. Chemical Reviews. 2022, 122(17): 13989-14017.
