Optimizing Transmembrane Protein Assemblies in Nanodiscs for Structural Studies: A Comprehensive Manual
Membrane protein structures offer a more accurate basis for understanding their functional correlates when derived from full-length proteins in their native lipid environment. Producing such samples has been a primary challenge in the field. Here, we present robust, step-by-step biochemical and biop...
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| Format: | Article |
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Bio-protocol LLC
2024-11-01
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| author | Fernando Vilela Cécile Sauvanet Armel Bezault Niels Volkmann Dorit Hanein |
| author_facet | Fernando Vilela Cécile Sauvanet Armel Bezault Niels Volkmann Dorit Hanein |
| author_sort | Fernando Vilela |
| collection | DOAJ |
| description | Membrane protein structures offer a more accurate basis for understanding their functional correlates when derived from full-length proteins in their native lipid environment. Producing such samples has been a primary challenge in the field. Here, we present robust, step-by-step biochemical and biophysical protocols for generating monodisperse assemblies of full-length transmembrane proteins within lipidic environments. These protocols are particularly tailored for cases where the size and molecular weight of the proteins align closely with those of the lipid islands (nanodiscs). While designed for single-span bitopic membrane proteins, these protocols can be easily extended to proteins with multiple transmembrane domains. The insights presented have broad implications across diverse fields, including biophysics, structural biology, and cryogenic electron microscopy (cryo-EM) studies. |
| format | Article |
| id | doaj-art-ee11f11fdea44e73b5f222f2a99d36f1 |
| institution | OA Journals |
| issn | 2331-8325 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | Bio-protocol LLC |
| record_format | Article |
| series | Bio-Protocol |
| spelling | doaj-art-ee11f11fdea44e73b5f222f2a99d36f12025-08-20T02:35:30ZengBio-protocol LLCBio-Protocol2331-83252024-11-01142110.21769/BioProtoc.5099Optimizing Transmembrane Protein Assemblies in Nanodiscs for Structural Studies: A Comprehensive ManualFernando Vilela0Cécile Sauvanet1Armel Bezault2Niels Volkmann3Dorit Hanein4Structural Studies of Macromolecular Machines in Cellulo Unit, Department of Structural Biology and Chemistry, Institut Pasteur, Université Paris Cité, CNRS UMR3528, Paris, FranceStructural Image Analysis Unit, Department of Structural Biology and Chemistry, Institut Pasteur, Université Paris Cité, CNRS UMR3528, Paris, FranceStructural Studies of Macromolecular Machines in Cellulo Unit, Department of Structural Biology and Chemistry, Institut Pasteur, Université Paris Cité, CNRS UMR3528, Paris, FranceStructural Image Analysis Unit, Department of Structural Biology and Chemistry, Institut Pasteur, Université Paris Cité, CNRS UMR3528, Paris, FranceStructural Studies of Macromolecular Machines in Cellulo Unit, Department of Structural Biology and Chemistry, Institut Pasteur, Université Paris Cité, CNRS UMR3528, Paris, FranceStructural Image Analysis Unit, Department of Structural Biology and Chemistry, Institut Pasteur, Université Paris Cité, CNRS UMR3528, Paris, FranceStructural Image Analysis Unit, Department of Structural Biology and Chemistry, Institut Pasteur, Université Paris Cité, CNRS UMR3528, Paris, FranceDepartment of Biological Engineering; Department of Electrical and Computer Engineering University of California, Santa Barbara, CA, USADepartment of Chemistry and Biochemistry, Department of Biological Engineering, University of California, Santa Barbara, CA, USAMembrane protein structures offer a more accurate basis for understanding their functional correlates when derived from full-length proteins in their native lipid environment. Producing such samples has been a primary challenge in the field. Here, we present robust, step-by-step biochemical and biophysical protocols for generating monodisperse assemblies of full-length transmembrane proteins within lipidic environments. These protocols are particularly tailored for cases where the size and molecular weight of the proteins align closely with those of the lipid islands (nanodiscs). While designed for single-span bitopic membrane proteins, these protocols can be easily extended to proteins with multiple transmembrane domains. The insights presented have broad implications across diverse fields, including biophysics, structural biology, and cryogenic electron microscopy (cryo-EM) studies.https://bio-protocol.org/en/bpdetail?id=5099&type=0 |
| spellingShingle | Fernando Vilela Cécile Sauvanet Armel Bezault Niels Volkmann Dorit Hanein Optimizing Transmembrane Protein Assemblies in Nanodiscs for Structural Studies: A Comprehensive Manual Bio-Protocol |
| title | Optimizing Transmembrane Protein Assemblies in Nanodiscs for Structural Studies: A Comprehensive Manual |
| title_full | Optimizing Transmembrane Protein Assemblies in Nanodiscs for Structural Studies: A Comprehensive Manual |
| title_fullStr | Optimizing Transmembrane Protein Assemblies in Nanodiscs for Structural Studies: A Comprehensive Manual |
| title_full_unstemmed | Optimizing Transmembrane Protein Assemblies in Nanodiscs for Structural Studies: A Comprehensive Manual |
| title_short | Optimizing Transmembrane Protein Assemblies in Nanodiscs for Structural Studies: A Comprehensive Manual |
| title_sort | optimizing transmembrane protein assemblies in nanodiscs for structural studies a comprehensive manual |
| url | https://bio-protocol.org/en/bpdetail?id=5099&type=0 |
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