An engineered thermal-shift screen reveals specific lipid preferences of eukaryotic and prokaryotic membrane proteins.

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Nji E, Chatzikyriakidou Y, Landreh M, Drew D

Nat Commun 9 (1) 4253 [2018-10-12; online 2018-10-12]

Membrane bilayers are made up of a myriad of different lipids that regulate the functional activity, stability, and oligomerization of many membrane proteins. Despite their importance, screening the structural and functional impact of lipid-protein interactions to identify specific lipid requirements remains a major challenge. Here, we use the FSEC-TS assay to show cardiolipin-dependent stabilization of the dimeric sodium/proton antiporter NhaA, demonstrating its ability to detect specific protein-lipid interactions. Based on the principle of FSEC-TS, we then engineer a simple thermal-shift assay (GFP-TS), which facilitates the high-throughput screening of lipid- and ligand- interactions with membrane proteins. By comparing the thermostability of medically relevant eukaryotic membrane proteins and a selection of bacterial counterparts, we reveal that eukaryotic proteins appear to have evolved to be more dependent to the presence of specific lipids.

PubMed 30315156

DOI 10.1038/s41467-018-06702-3

Crossref 10.1038/s41467-018-06702-3

pmc: PMC6185904
pii: 10.1038/s41467-018-06702-3