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Defining state-selective lipid binding to brain GPCRs - introducing REVEAL

Lawrence, S. A. S.; Kirschbaum, C.; Lutomski, C. A.; Song, H.; Hinkle, J. D.; Butroid, F. I.; Melani, R.; Syka, J. E. P.; Mullen, C.; El-Baba, T. J.; Robinson, C. V.

2026-05-05 biophysics
10.64898/2026.05.01.722320 bioRxiv
Show abstract

Membrane lipids are central regulators of G protein-coupled receptor (GPCR) function. Defining receptor-specific lipid interactions in native, fully modified mammalian systems remains challenging. Extensive post-translational modifications generate heterogeneous proteoforms that confound conventional mass spectrometry approaches. Here we introduce REVEAL (REceptor enVironment Elucidation by Activated Lipid-release), an automated native top-down mass spectrometry strategy that discriminates specifically bound lipids from background. Applied here to intact, heterogeneous mammalian membrane protein complexes, incubated with a brain polar lipid extract (>1000 components), we define receptor-specific lipid-binding for two neuronal class C GPCRs. We show that agonism remodels lipid occupancy, selectively enriching a reduced repertoire of bound lipids. Plasmalogen lipids emerge as persistent binders across all conformational states of the metabotropic glycine receptor and are preferentially depleted under oxidative stress, implying a protective role at the receptor surface. These findings position lipids as dynamic regulators of both function and response to the cellular redox environment.

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