The Membrane Proximal Domain of TRPV1 and TRPV2 Channels Mediates Protein–Protein Interactions and Lipid Binding In Vitro

Biophysics Unit, Department of Biochemistry and Molecular Biology, School of Medicine, Universitat Autònoma de Barcelona, 08193 Cerdanyola del Vallés, Catalonia, Spain

Departament d’Enginyeria Agroalimentària i Biotecnologia, Universitat Politècnica de Catalunya, 08860 Barcelona, Catalonia, Spain

Institut de Biologie Intégrative de la Cellule, CEA-Saclay, 91191 Gif-sur-Yvette, France

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Institut des Sciences du Vivant Frédéric-JOLIOT, CEA-Saclay, 91191 Gif-sur-Yvette, France

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Institut de Neurociències, Universitat Autònoma de Barcelona, 08193 Cerdanyola del Vallés, Catalonia, Spain

Author to whom correspondence should be addressed.

Int. J. Mol. Sci. 2019, 20(3), 682; https://doi.org/10.3390/ijms20030682

Received: 14 January 2019 / Revised: 30 January 2019 / Accepted: 31 January 2019 / Published: 5 February 2019

(This article belongs to the Special Issue Membrane Channels in Human Diseases: The Spanish Ion Channel Initiative Consortium (SICI))

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Abstract

Constitutive or regulated membrane protein trafficking is a key cell biology process. Transient receptor potential channels are somatosensory proteins in charge of detecting several physical and chemical stimuli, thus requiring fine vesicular trafficking. The membrane proximal or pre-S1 domain (MPD) is a highly conserved domain in transient receptor potential channels from the vanilloid (TRPV) subfamily. MPD shows traits corresponding to protein-protein and lipid-protein interactions, and protein regulatory regions. We have expressed MPD of TRPV1 and TRPV2 as green fluorescente protein (GFP)-fusion proteins to perform an in vitro biochemical and biophysical characterization. Pull-down experiments indicate that MPD recognizes and binds Soluble N-ethylmaleimide-sensitive factor Attachment Protein Receptors (SNARE). Synchrotron radiation scattering experiments show that this domain does not self-oligomerize. MPD interacts with phosphatidic acid (PA), a metabolite of the phospholipase D (PLD) pathway, in a specific manner as shown by lipid strips and Trp fluorescence quenching experiments. We show for the first time, to the best of our knowledge, the binding to PA of an N-terminus domain in TRPV channels. The presence of a PA binding domain in TRPV channels argues for putative PLD regulation. Findings in this study open new perspectives to understand the regulated and constitutive trafficking of TRPV channels exerted by protein-protein and lipid-protein interactions. View Full-Text

Keywords: Transient Receptor Potential (TRP) channels; exocytosis; biophysics; protein–protein interactions; lipid-protein interactions

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MDPI and ACS Style

Doñate-Macián, P.; Álvarez-Marimon, E.; Sepulcre, F.; Vázquez-Ibar, J.L.; Perálvarez-Marín, A. The Membrane Proximal Domain of TRPV1 and TRPV2 Channels Mediates Protein–Protein Interactions and Lipid Binding In Vitro. Int. J. Mol. Sci. 2019, 20, 682. https://doi.org/10.3390/ijms20030682

AMA Style

Doñate-Macián P, Álvarez-Marimon E, Sepulcre F, Vázquez-Ibar JL, Perálvarez-Marín A. The Membrane Proximal Domain of TRPV1 and TRPV2 Channels Mediates Protein–Protein Interactions and Lipid Binding In Vitro. International Journal of Molecular Sciences. 2019; 20(3):682. https://doi.org/10.3390/ijms20030682

Chicago/Turabian Style

Doñate-Macián, Pau, Elena Álvarez-Marimon, Francesc Sepulcre, José Luis Vázquez-Ibar, and Alex Perálvarez-Marín. 2019. "The Membrane Proximal Domain of TRPV1 and TRPV2 Channels Mediates Protein–Protein Interactions and Lipid Binding In Vitro" International Journal of Molecular Sciences 20, no. 3: 682. https://doi.org/10.3390/ijms20030682

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The Membrane Proximal Domain of TRPV1 and TRPV2 Channels Mediates Protein–Protein Interactions and Lipid Binding In Vitro

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