The membrane proximal domain of TRPV1 and TRPV2 channels mediates protein–protein interactions and lipid binding in vitro

Pau Doñate-Macián, Elena Álvarez-Marimon, Francesc Sepulcre, José Luis Vázquez-Ibar, Alex Perálvarez-Marín

Research output: Contribution to journalArticleResearch

1 Citation (Scopus)

Abstract

© 2019 by the authors. Licensee MDPI, Basel, Switzerland. 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.
Original languageEnglish
Article number682
JournalInternational Journal of Molecular Sciences
Volume20
DOIs
Publication statusPublished - 1 Jan 2019

Keywords

  • Biophysics
  • Exocytosis
  • Lipid-protein interactions
  • Protein–protein interactions
  • Transient Receptor Potential (TRP) channels

Fingerprint Dive into the research topics of 'The membrane proximal domain of TRPV1 and TRPV2 channels mediates protein–protein interactions and lipid binding in vitro'. Together they form a unique fingerprint.

Cite this