New Protocol for Screening Membrane Protein Stability
Six research groups, among them three from CSSB, have developed a protocol that will simplify the process of solubilising integral membrane proteins (IMPs), as they reported in Scientific Reports. The development of the protocol is the first result of a collaboration between various research groups using the Protein Characterisation (PC) facility at CSSB.
“Working with membrane proteins is challenging,” says corresponding author María García Alai, the EMBL group leader responsible for running the PC facility. “While they’re in the membrane, they’re not soluble. So you have to get them out, which is the most difficult part. When you do that, you might be altering the structure of the proteins, so you never know exactly what is happening.”
The first step of working with IMPs is to extract them from the membrane using detergents. This can alter the protein, which then has to be purified and reintegrated into a membrane-like environment. Experimenting with different detergents to get the best result is expensive, because the detergent has to be removed in various technical steps. García’s team and their collaborators showed that it’s possible to measure the stability and solubility of IMPs by diluting them into different detergents. The result was a protocol that allows the identification of suitable conditions for membrane proteins during purification. “This will be very useful for all the laboratories working with membrane proteins. We show a lot of evidence that this pipeline is the way to go,” says García.
The Protein Characterisation facility is one of five multi-user facilities at CSSB. It supports users with the design, execution and data analysis of biophysical experiments aimed at the characterization of proteins, protein complexes and interactions between proteins and other types of molecules. “One of the most important things about the CSSB concept is that we’re not only doing research, but also contributing with facilities for the scientific community,” says García Alai. “All the different scientific institutions that are part of CSSB work together and push forward the infrastructure, facilities and scientific services. And this is the first result.”
Original EMBL Article:
García Alai et al. High-throughput stability screening for detergent-solubilized membrane proteins, Scientific Reports. Published online 17 July 2019. DOI: https://doi.org/10.1038/s41598-019-46686-8