CETSA® on Membrane Proteins
CETSA® has primarily been applied to soluble proteins which, upon heating, denaturate to form insoluble aggregates by exposing hydrophobic surfaces. Nevertheless, CETSA® has successfully been applied to a number of membrane-associated proteins including GPCRs and transporters.
Case studies on how to deploy CETSA® on insoluble protein targets such as membrane proteins have been published, for example by Kawatkar et al. in 2019. This study describes the thorough experimental work undertaken to develop assay protocols for the more challenging multi-pass transmembrane proteins. This is hugely valuable to drug discovery researchers in that by demonstrating how CETSA® can be used for membrane-bound proteins, such as ion channels and transporters, scientists now have an alternative to biochemical or biophysical methods.
In a recent study by Kalxdorf et al., CETSA® Explore was combined with selective enrichment of glycosylated cell surface proteins. Included in the publication is the profiling of a range of small molecule compounds and biological stimuli which demonstrated the potential of the method to identify and study membrane–bound protein targets.
Kawatkar A, Schefter M, Hermansson NO, Snijder A, et al. CETSA beyond Soluble Targets: a Broad Application to Multipass Transmembrane Proteins. (2019) ACS Chem Biol. DOI: 10.1021/acschembio.9b00399
Kalxdorf M, Günthner I, et al. Cell surface thermal proteome profiling tracks perturbations and drug targets on the plasma membrane. (2021) Nature Methods. DOI: 10.1038/s41592-020-01