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Article Dans Une Revue Cell Reports Methods Année : 2023

B2LiVe, a label-free 1D-NMR method to quantify the binding of amphitropic peptides or proteins to membrane vesicles

Résumé

Amphitropic proteins and peptides reversibly partition from solution to membrane, a key process that regulates their functions. Experimental approaches classically used to measure protein partitioning into lipid bilayers, such as fluorescence and circular dichroism, are hardly usable when the peptides or proteins do not exhibit significant polarity and/or conformational changes upon membrane binding. Here, we describe binding to lipid vesicles (B2LiVe), a simple, robust, and widely applicable nuclear magnetic resonance (NMR) method to determine the solution-to-membrane partitioning of unlabeled proteins or peptides. B2LiVe relies on previously described proton 1D-NMR fast-pulsing techniques. Membrane partitioning induces a large line broadening, leading to a loss of protein signals; therefore, the decrease of the NMR signal directly measures the fraction of membrane-bound protein. The method uses low polypeptide concentrations and has been validated on several membrane-interacting polypeptides, ranging from 3 to 54 kDa, with membrane vesicles of different sizes and various lipid compositions.
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Origine : Publication financée par une institution
Licence : CC BY NC ND - Paternité - Pas d'utilisation commerciale - Pas de modification

Dates et versions

pasteur-04356880 , version 1 (02-01-2023)
pasteur-04356880 , version 2 (20-12-2023)

Licence

Paternité - Pas d'utilisation commerciale - Pas de modification

Identifiants

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Mirko Sadi, Nicolas Carvalho, Corentin Léger, Bruno Vitorge, Daniel Ladant, et al.. B2LiVe, a label-free 1D-NMR method to quantify the binding of amphitropic peptides or proteins to membrane vesicles. Cell Reports Methods, 2023, 3 (11), pp.100624. ⟨10.1016/j.crmeth.2023.100624⟩. ⟨pasteur-04356880v2⟩

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