Back to Search Start Over

Expression, Biochemistry, and Stabilization with Camel Antibodies of Membrane Proteins: Case Study of the Mouse 5-HT3 Receptor

Authors :
Aline Desmyter
Horst Vogel
Lamia Mebarki
Takashi Tomizaki
Ruud Hovius
Romain Wyss
Sonja Minniberger
Xiao-Dan Li
Christophe Moreau
Lucie Peclinovska
Ghérici Hassaine
Cédric Deluz
Hugues Nury
Luigino Grasso
Henning Stahlberg
Laboratory of Physical Chemistry of Polymers and Membranes
Ecole Polytechnique Fédérale de Lausanne (EPFL)
Biozentrum
University of Basel (Unibas)
Architecture et fonction des macromolécules biologiques (AFMB)
Institut National de la Recherche Agronomique (INRA)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)
Institut de biologie structurale (IBS - UMR 5075)
Institut de Recherche Interdisciplinaire de Grenoble (IRIG)
Direction de Recherche Fondamentale (CEA) (DRF (CEA))
Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA))
Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA)-Centre National de la Recherche Scientifique (CNRS)
Laboratory of Biomolecular Research
Paul Scherrer Institute (PSI)
Biozentrum [Basel, Suisse]
Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU)-Institut National de la Recherche Agronomique (INRA)
Institut de biologie structurale (IBS - UMR 5075 )
Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG)
Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)
Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG)
Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)
Lacapere, Jean-Jacques
Source :
Methods in Molecular Biology, Methods in Molecular Biology, Humana Press/Springer Imprint, 2018, pp.139-168. ⟨10.1007/978-1-4939-7151-0_8⟩, Methods in Molecular Biology, 2018, pp.139-168. ⟨10.1007/978-1-4939-7151-0_8⟩, Methods in Molecular Biology ISBN: 9781493971497, Methods in Molecular Biology-Membrane Protein Structure and Function Characterization, Methods in Molecular Biology, Membrane Protein Structure and Function Characterization
Publication Year :
2018
Publisher :
HAL CCSD, 2018.

Abstract

International audience; There is growing interest in the use of mammalian protein expression systems, and in the use of antibody-derived chaperones, for structural studies. Here, we describe protocols ranging from the production of recombinant membrane proteins in stable inducible cell lines to biophysical characterization of purified membrane proteins in complex with llama antibody domains. These protocols were used to solve the structure of the mouse 5-HT3 serotonin receptor but are of broad applicability for crystallization or cryo-electron microscopy projects.

Subjects

Subjects :
0301 basic medicine
[SDV.BIO]Life Sciences [q-bio]/Biotechnology
[SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology
Gene Expression
Crystallography, X-Ray
[SDV.IMM.II]Life Sciences [q-bio]/Immunology/Innate immunity
law.invention
MESH: Recombinant Proteins
Mice
law
[SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases
MESH: Animals
Integral membrane protein
[SDV.MHEP.ME]Life Sciences [q-bio]/Human health and pathology/Emerging diseases
biology
[SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM]
Protein Stability
Peripheral membrane protein
MESH: Camelus
[SDV.BIBS]Life Sciences [q-bio]/Quantitative Methods [q-bio.QM]
Recombinant Proteins
[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM]
[SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM]
Biochemistry
[SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology
Recombinant DNA
MESH: Membrane Proteins
MESH: Cryoelectron Microscopy
Antibody
Camelus
MESH: Gene Expression
MESH: Receptors, Serotonin, 5-HT3
Cys-loop receptor
VHH
5-HT3 receptor
Antibodies
Cell Line
03 medical and health sciences
MESH: Protein Stability
Animals
Humans
[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM]
MESH: Mice
5-HT receptor
MESH: Humans
MESH: Antibodies
Cryoelectron Microscopy
Membrane Proteins
MESH: Crystallography, X-Ray
[SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology
Lama antibody
MESH: Cell Line
030104 developmental biology
Membrane protein
Cell culture
biology.protein
[SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology
Receptors, Serotonin, 5-HT3
Stable cell line

Details

Language :
English
ISBN :
978-1-4939-7149-7
978-1-4939-7151-0
ISSN :
10643745 and 19406029
ISBNs :
9781493971497 and 9781493971510
Database :
OpenAIRE
Journal :
Methods in Molecular Biology, Methods in Molecular Biology, Humana Press/Springer Imprint, 2018, pp.139-168. ⟨10.1007/978-1-4939-7151-0_8⟩, Methods in Molecular Biology, 2018, pp.139-168. ⟨10.1007/978-1-4939-7151-0_8⟩, Methods in Molecular Biology ISBN: 9781493971497, Methods in Molecular Biology-Membrane Protein Structure and Function Characterization, Methods in Molecular Biology, Membrane Protein Structure and Function Characterization
Accession number :
edsair.doi.dedup.....b97109ac8dfda1b9855a018b3376074a
Full Text :
https://doi.org/10.1007/978-1-4939-7151-0_8⟩