Abstract
Caveolins, major components of small plasma membrane invaginations called caveolae, play a role in signaling, particularly in mechanosignaling. These proteins are known to interact with a variety of effector molecules, including G-protein-coupled receptors, Src family kinases, ion channels, endothelial nitric oxide synthase (eNOS), adenylyl cyclases, protein kinase A (PKA), and mitogen-activated PKs (MAPKs). There is, however, speculation on the relevance of these interactions and the mechanisms by which caveolins may control intracellular signaling. This chapter introduces a method of isolation of giant plasma membrane-derived vesicles (GPMVs), which possess full complexity of membrane they originate from, thus comprising an excellent platform to revisit some of the previously described interactions in a cleaner environment and possibly identifying new binding partners. It is also a powerful technique for studying membrane mechanics, as it was previously used to demonstrate the role of caveolae in mechanoprotection.
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Acknowledgments
The authors would like to thank Christophe Lamaze and Patricia Bassereau and all the people from the Membrane mechanics and dynamics of intracellular signaling laboratory and Membranes and Cellular Functions laboratory. The facilities as well as scientific and technical assistance from staff in the PICT-IBiSA/Nikon Imaging Centre at Institut Curie-CNRS and the France-BioImaging infrastructure (No. ANR-10-INSB-04) are acknowledged. This work was supported by institutional grants from the Curie Institute, INSERM, and CNRS, and by specific grants from Association Française contre les Myopathies (AFM): 22337 (to J.P.) and CAV-STRESS-MUS (14266 to C.M.B.). J.P. was funded by Polish Ministry of Science and Higher Education Mobility Plus program (1668/MOB/V/2017/0) and Labex CelTisPhyBio. The Lamaze team, the PICT-IBiSA/Nikon Imaging Centre at Institut Curie-CNRS, and the France-BioImaging infrastructure are members of Labex CelTisPhyBio (No. ANR-10-LBX-0038) and of IDEX PSL (No. ANR-10-IDEX-0001-02 PSL).
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Podkalicka, J., Blouin, C.M. (2020). GPMVs as a Tool to Study Caveolin-Interacting Partners. In: Blouin, C. (eds) Caveolae. Methods in Molecular Biology, vol 2169. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0732-9_8
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DOI: https://doi.org/10.1007/978-1-0716-0732-9_8
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