B2LiVe, a label-free 1D-NMR method to quantify the binding of amphitropic peptides or proteins to membrane vesicles
Summary: 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 t...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-11-01
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| Series: | Cell Reports: Methods |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2667237523002886 |
| _version_ | 1797526197754658816 |
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| author | Mirko Sadi Nicolas Carvalho Corentin Léger Bruno Vitorge Daniel Ladant J. Iñaki Guijarro Alexandre Chenal |
| author_facet | Mirko Sadi Nicolas Carvalho Corentin Léger Bruno Vitorge Daniel Ladant J. Iñaki Guijarro Alexandre Chenal |
| author_sort | Mirko Sadi |
| collection | DOAJ |
| description | Summary: 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. Motivation: Characterization of the interaction of peptides and proteins with lipid membranes is relevant for the study of various biological processes and is often challenging for polypeptides, which do not possess intrinsic fluorophores and do not exhibit significant structural content changes, as well as for those characterized by low membrane affinities. To meet these challenges, we have developed a label-free 1D-1H-NMR-based experimental approach, named B2LiVe, to measure the binding of polypeptides to lipid vesicles. B2LiVe complements the arsenal of label-free biophysical assays available to characterize protein-membrane interactions. |
| first_indexed | 2024-03-10T09:25:50Z |
| format | Article |
| id | doaj.art-98dbe9fa67214533aca3fb57a878a923 |
| institution | Directory Open Access Journal |
| issn | 2667-2375 |
| language | English |
| last_indexed | 2024-03-10T09:25:50Z |
| publishDate | 2023-11-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Cell Reports: Methods |
| spelling | doaj.art-98dbe9fa67214533aca3fb57a878a9232023-11-22T04:49:25ZengElsevierCell Reports: Methods2667-23752023-11-01311100624B2LiVe, a label-free 1D-NMR method to quantify the binding of amphitropic peptides or proteins to membrane vesiclesMirko Sadi0Nicolas Carvalho1Corentin Léger2Bruno Vitorge3Daniel Ladant4J. Iñaki Guijarro5Alexandre Chenal6Institut Pasteur, Université de Paris Cité, CNRS UMR3528, Biochemistry of Macromolecular Interactions Unit, 75015 Paris, France; Université de Paris Cité, 75005 Paris, FranceInstitut Pasteur, Université de Paris Cité, CNRS UMR3528, Biochemistry of Macromolecular Interactions Unit, 75015 Paris, France; Université de Paris Cité, 75005 Paris, FranceInstitut Pasteur, Université de Paris Cité, CNRS UMR3528, Biochemistry of Macromolecular Interactions Unit, 75015 Paris, FranceInstitut Pasteur, Université de Paris Cité, CNRS UMR3528, Biological NMR and HDX-MS Technological Platform, 75015 Paris, FranceInstitut Pasteur, Université de Paris Cité, CNRS UMR3528, Biochemistry of Macromolecular Interactions Unit, 75015 Paris, FranceInstitut Pasteur, Université de Paris Cité, CNRS UMR3528, Biological NMR and HDX-MS Technological Platform, 75015 Paris, France; Corresponding authorInstitut Pasteur, Université de Paris Cité, CNRS UMR3528, Biochemistry of Macromolecular Interactions Unit, 75015 Paris, France; Corresponding authorSummary: 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. Motivation: Characterization of the interaction of peptides and proteins with lipid membranes is relevant for the study of various biological processes and is often challenging for polypeptides, which do not possess intrinsic fluorophores and do not exhibit significant structural content changes, as well as for those characterized by low membrane affinities. To meet these challenges, we have developed a label-free 1D-1H-NMR-based experimental approach, named B2LiVe, to measure the binding of polypeptides to lipid vesicles. B2LiVe complements the arsenal of label-free biophysical assays available to characterize protein-membrane interactions.http://www.sciencedirect.com/science/article/pii/S2667237523002886CP: Molecular biologyCP: Cell biology |
| spellingShingle | Mirko Sadi Nicolas Carvalho Corentin Léger Bruno Vitorge Daniel Ladant J. Iñaki Guijarro Alexandre Chenal B2LiVe, a label-free 1D-NMR method to quantify the binding of amphitropic peptides or proteins to membrane vesicles Cell Reports: Methods CP: Molecular biology CP: Cell biology |
| title | B2LiVe, a label-free 1D-NMR method to quantify the binding of amphitropic peptides or proteins to membrane vesicles |
| title_full | B2LiVe, a label-free 1D-NMR method to quantify the binding of amphitropic peptides or proteins to membrane vesicles |
| title_fullStr | B2LiVe, a label-free 1D-NMR method to quantify the binding of amphitropic peptides or proteins to membrane vesicles |
| title_full_unstemmed | B2LiVe, a label-free 1D-NMR method to quantify the binding of amphitropic peptides or proteins to membrane vesicles |
| title_short | B2LiVe, a label-free 1D-NMR method to quantify the binding of amphitropic peptides or proteins to membrane vesicles |
| title_sort | b2live a label free 1d nmr method to quantify the binding of amphitropic peptides or proteins to membrane vesicles |
| topic | CP: Molecular biology CP: Cell biology |
| url | http://www.sciencedirect.com/science/article/pii/S2667237523002886 |
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