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Article Dans Une Revue Nature Methods Année : 2023

Direct determination of oligomeric organization of integral membrane proteins and lipids from intact customizable bilayer

Aniruddha Panda
Anna Duncan
Alexander Welch
Sathish Ramakrishnan
Frédéric Pincet
Lan Guan
Shyam Krishnakumar
Kallol Gupta

Résumé

Hierarchical organization of integral membrane proteins (IMP) and lipids at the membrane is essential for regulating myriad downstream signaling. A quantitative understanding of these processes requires both detections of oligomeric organization of IMPs and lipids directly from intact membranes and determination of key membrane components/properties that regulate them. Addressing this, we have developed a platform that enables native mass spectrometry (nMS) analysis of IMP-lipid complexes directly from intact and customizable lipid membranes. Both the lipid composition and membrane properties (such as curvature, tension, fluidity) of these bilayers can be precisely customized to a target membrane. Subsequent direct nMS analysis of these intact proteo-lipid vesicles can yield the oligomeric states of the embedded IMPs, identify bound lipids, and determine the membrane properties that can regulate the observed IMP-lipid organization. Applying this, we show how lipid binding regulates neurotransmitter release and how membrane composition regulates the functional oligomeric state of a transporter.
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Dates et versions

hal-04161556 , version 1 (13-07-2023)

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Aniruddha Panda, Fabian Giska, Anna Duncan, Alexander Welch, Caroline Brown, et al.. Direct determination of oligomeric organization of integral membrane proteins and lipids from intact customizable bilayer. Nature Methods, 2023, 20 (6), pp.891-897. ⟨10.1038/s41592-023-01864-5⟩. ⟨hal-04161556⟩
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