9f0c77629ceb764274e3f81e149980e6 pmc_30764505.pdf 191ae6a89a3db97f7849045b3d99c154f2820733 pmc_30764505.pdf 8cc540a885452bfaabebff546fdda420a0e132b80d96cf3490e2ce7be2239579 pmc_30764505.pdf Title: The Membrane Proximal Domain of TRPV1 and TRPV2 Channels Mediates Protein–Protein Interactions and Lipid Binding In Vitro Subject: 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. Keywords: Transient Receptor Potential (TRP) channels; exocytosis; biophysics; protein–protein interactions; lipid-protein interactions Author: Pau Doñate-Macián, Elena Álvarez-Marimon, Francesc Sepulcre, José Luis Vázquez-Ibar and Alex Perálvarez-Marín Creator: LaTeX with hyperref package Producer: pdfTeX-1.40.17 CreationDate: Tue Feb 5 12:12:56 2019 CET ModDate: Tue Feb 5 12:12:56 2019 CET Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 11 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 2277626 bytes Optimized: no PDF version: 1.5 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- KRBWEG+URWPalladioL-Ital Type 1 Custom yes yes no 100 0 WMLZPF+URWPalladioL-Bold Type 1 Custom yes yes no 101 0 ILUGHX+VnURWPalladioL-Bold Type 1 Custom yes yes no 102 0 MVNLKZ+URWPalladioL-Roma Type 1 Custom yes yes no 103 0 SXQPWE+VnURWPalladioL Type 1 Custom yes yes no 104 0 CKUPDE+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 109 0 IRWHZX+TimesNewRomanPSMT Type 1C WinAnsi yes yes no 110 0 MCBCCI+SourceSansRoman-Regular Type 1C Custom yes yes no 129 0 MCBCCM+SourceSansRoman.613wght Type 1C Custom yes yes no 130 0 MCBCCO+SourceSansRoman-Bold Type 1C Custom yes yes no 131 0 MPUDFL+PazoMath Type 1 Builtin yes yes no 166 0 GGRFCO+CMSY10 Type 1 Builtin yes yes no 201 0 CAMXTK+EURM10 Type 1 Builtin yes yes no 203 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2020-11-19 06:17:25 CET RepresentationInformation: pmc_30764505.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2020-08-03 03:44:54 CEST Size: 2277626 Format: PDF Version: 1.5 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 0 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: ViewerPreferences: HideToolbar: false HideMenubar: false HideWindowUI: false FitWindow: true CenterWindow: false DisplayDocTitle: false NonFullScreenPageMode: UseNone Direction: L2R ViewArea: CropBox ViewClip: CropBox PrintArea: CropBox PageClip: CropBox PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: ぉのぴひはつふっぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳散瑩潮⸲ Item: Title: げづびふぬぴびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㈮ㄩ‾㸊⡜㌷㙜㌷㝜〰ごげぐざびぜ〴ぜ〰ちのつぜ〴ぜ〰こぎぁげぅ〭ぐひはぴづどのぜ〴ぜ〰きぐい〭きづつどちぴづつぜ〴ぜ〰ぉのぴづひちっぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㈮㈩‾㸊⡜㌷㙜㌷㝜〰きぐい〭きづつどちぴづつぜ〴ぜ〰がどばどつぜ〴ぜ〰あどのつどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㈮㌩‾㸊⡜㌷㙜㌷㝜〰きぐいぜ〴ぜ〰くぬどではねづひどぺちぴどはのぜ〴ぜ〰こぴちぴづぜ〴〩਼㰠⽓ 䝯呯 䐠⡳散瑩潮⸳ Children: Item: Title: ごげぐざびぜ〴ぜ〰ちのつぜ〴ぜ〰こぎぁげぅ〭ぐひはぴづどのぜ〴ぜ〰きぐい〭きづつどちぴづつぜ〴ぜ〰ぉのぴづひちっぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㈮㈩‾㸊⡜㌷㙜㌷㝜〰きぐい〭きづつどちぴづつぜ〴ぜ〰がどばどつぜ〴ぜ〰あどのつどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㈮㌩‾㸊⡜㌷㙜㌷㝜〰きぐいぜ〴ぜ〰くぬどではねづひどぺちぴどはのぜ〴ぜ〰こぴちぴづぜ〴〩਼㰠⽓ 䝯呯 䐠⡳散瑩潮⸳ Item: Title: きぐい〭きづつどちぴづつぜ〴ぜ〰がどばどつぜ〴ぜ〰あどのつどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㈮㌩‾㸊⡜㌷㙜㌷㝜〰きぐいぜ〴ぜ〰くぬどではねづひどぺちぴどはのぜ〴ぜ〰こぴちぴづぜ〴〩਼㰠⽓ 䝯呯 䐠⡳散瑩潮⸳ Item: Title: きぐいぜ〴ぜ〰くぬどではねづひどぺちぴどはのぜ〴ぜ〰こぴちぴづぜ〴〩਼㰠⽓ 䝯呯 䐠⡳散瑩潮⸳ Item: Title: いどびっふびびどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳散瑩潮⸴ Item: Title: きちぴづひどちぬびぜ〴ぜ〰ちのつぜ〴ぜ〰きづぴとはつびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮ㄩ‾㸊⡜㌷㙜㌷㝜〰いぎぁぜ〴ぜ〰ぐぬちびねどつびぜ〴ぜ〰ちのつぜ〴ぜ〰ぃぬはのどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㈩‾㸊⡜㌷㙜㌷㝜〰げづっはねぢどのちのぴぜ〴ぜ〰ぐひはぴづどのぜ〴ぜ〰ぅへばひづびびどはのぜ〴ぜ〰ちのつぜ〴ぜ〰ぐふひどてどっちぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㌩‾㸊⡜㌷㙜㌷㝜〰ぃづぬぬぜ〴ぜ〰ぃふぬぴふひづびぜ〴ぜ〰ちのつぜ〴ぜ〰ごひちのびてづっぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㐩‾㸊⡜㌷㙜㌷㝜〰ぉねねふのはぢぬはぴぴどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㔩‾㸊⡜㌷㙜㌷㝜〰ぃは〭ぉねねふのはばひづっどばどぴちぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㘩‾㸊⡜㌷㙜㌷㝜〰ぐふぬぬぜ〴ぜ〰いはぷのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㜩‾㸊⡜㌷㙜㌷㝜〰がどばどつぜ〴ぜ〰こぴひどばびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㠩‾㸊⡜㌷㙜㌷㝜〰ごひべばぴはばとちのぜ〴ぜ〰けふづのっとどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㤩‾㸊⡜㌷㙜㌷㝜〰こぁじこぜ〴ぜ〰いちぴちぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㄰ Children: Item: Title: いぎぁぜ〴ぜ〰ぐぬちびねどつびぜ〴ぜ〰ちのつぜ〴ぜ〰ぃぬはのどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㈩‾㸊⡜㌷㙜㌷㝜〰げづっはねぢどのちのぴぜ〴ぜ〰ぐひはぴづどのぜ〴ぜ〰ぅへばひづびびどはのぜ〴ぜ〰ちのつぜ〴ぜ〰ぐふひどてどっちぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㌩‾㸊⡜㌷㙜㌷㝜〰ぃづぬぬぜ〴ぜ〰ぃふぬぴふひづびぜ〴ぜ〰ちのつぜ〴ぜ〰ごひちのびてづっぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㐩‾㸊⡜㌷㙜㌷㝜〰ぉねねふのはぢぬはぴぴどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㔩‾㸊⡜㌷㙜㌷㝜〰ぃは〭ぉねねふのはばひづっどばどぴちぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㘩‾㸊⡜㌷㙜㌷㝜〰ぐふぬぬぜ〴ぜ〰いはぷのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㜩‾㸊⡜㌷㙜㌷㝜〰がどばどつぜ〴ぜ〰こぴひどばびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㠩‾㸊⡜㌷㙜㌷㝜〰ごひべばぴはばとちのぜ〴ぜ〰けふづのっとどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㤩‾㸊⡜㌷㙜㌷㝜〰こぁじこぜ〴ぜ〰いちぴちぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㄰ Item: Title: げづっはねぢどのちのぴぜ〴ぜ〰ぐひはぴづどのぜ〴ぜ〰ぅへばひづびびどはのぜ〴ぜ〰ちのつぜ〴ぜ〰ぐふひどてどっちぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㌩‾㸊⡜㌷㙜㌷㝜〰ぃづぬぬぜ〴ぜ〰ぃふぬぴふひづびぜ〴ぜ〰ちのつぜ〴ぜ〰ごひちのびてづっぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㐩‾㸊⡜㌷㙜㌷㝜〰ぉねねふのはぢぬはぴぴどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㔩‾㸊⡜㌷㙜㌷㝜〰ぃは〭ぉねねふのはばひづっどばどぴちぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㘩‾㸊⡜㌷㙜㌷㝜〰ぐふぬぬぜ〴ぜ〰いはぷのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㜩‾㸊⡜㌷㙜㌷㝜〰がどばどつぜ〴ぜ〰こぴひどばびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㠩‾㸊⡜㌷㙜㌷㝜〰ごひべばぴはばとちのぜ〴ぜ〰けふづのっとどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㤩‾㸊⡜㌷㙜㌷㝜〰こぁじこぜ〴ぜ〰いちぴちぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㄰ Item: Title: ぃづぬぬぜ〴ぜ〰ぃふぬぴふひづびぜ〴ぜ〰ちのつぜ〴ぜ〰ごひちのびてづっぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㐩‾㸊⡜㌷㙜㌷㝜〰ぉねねふのはぢぬはぴぴどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㔩‾㸊⡜㌷㙜㌷㝜〰ぃは〭ぉねねふのはばひづっどばどぴちぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㘩‾㸊⡜㌷㙜㌷㝜〰ぐふぬぬぜ〴ぜ〰いはぷのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㜩‾㸊⡜㌷㙜㌷㝜〰がどばどつぜ〴ぜ〰こぴひどばびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㠩‾㸊⡜㌷㙜㌷㝜〰ごひべばぴはばとちのぜ〴ぜ〰けふづのっとどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㤩‾㸊⡜㌷㙜㌷㝜〰こぁじこぜ〴ぜ〰いちぴちぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㄰ Item: Title: ぉねねふのはぢぬはぴぴどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㔩‾㸊⡜㌷㙜㌷㝜〰ぃは〭ぉねねふのはばひづっどばどぴちぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㘩‾㸊⡜㌷㙜㌷㝜〰ぐふぬぬぜ〴ぜ〰いはぷのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㜩‾㸊⡜㌷㙜㌷㝜〰がどばどつぜ〴ぜ〰こぴひどばびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㠩‾㸊⡜㌷㙜㌷㝜〰ごひべばぴはばとちのぜ〴ぜ〰けふづのっとどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㤩‾㸊⡜㌷㙜㌷㝜〰こぁじこぜ〴ぜ〰いちぴちぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㄰ Item: Title: ぃは〭ぉねねふのはばひづっどばどぴちぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㘩‾㸊⡜㌷㙜㌷㝜〰ぐふぬぬぜ〴ぜ〰いはぷのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㜩‾㸊⡜㌷㙜㌷㝜〰がどばどつぜ〴ぜ〰こぴひどばびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㠩‾㸊⡜㌷㙜㌷㝜〰ごひべばぴはばとちのぜ〴ぜ〰けふづのっとどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㤩‾㸊⡜㌷㙜㌷㝜〰こぁじこぜ〴ぜ〰いちぴちぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㄰ Item: Title: ぐふぬぬぜ〴ぜ〰いはぷのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㜩‾㸊⡜㌷㙜㌷㝜〰がどばどつぜ〴ぜ〰こぴひどばびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㠩‾㸊⡜㌷㙜㌷㝜〰ごひべばぴはばとちのぜ〴ぜ〰けふづのっとどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㤩‾㸊⡜㌷㙜㌷㝜〰こぁじこぜ〴ぜ〰いちぴちぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㄰ Item: Title: がどばどつぜ〴ぜ〰こぴひどばびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㠩‾㸊⡜㌷㙜㌷㝜〰ごひべばぴはばとちのぜ〴ぜ〰けふづのっとどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㤩‾㸊⡜㌷㙜㌷㝜〰こぁじこぜ〴ぜ〰いちぴちぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㄰ Item: Title: ごひべばぴはばとちのぜ〴ぜ〰けふづのっとどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㤩‾㸊⡜㌷㙜㌷㝜〰こぁじこぜ〴ぜ〰いちぴちぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㄰ Item: Title: こぁじこぜ〴ぜ〰いちぴちぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㄰ Item: Title: きこぁぜ〴ぜ〰ちのつぜ〴ぜ〰ごひどつどねづのびどはのちぬぜ〴ぜ〰げづばひづびづのぴちぴどはのびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㄰ Item: Title: げづてづひづのっづび Filters: FilterPipeline: FlateDecode Fonts: Type1: Font: BaseFont: KRBWEG+URWPalladioL-Ital FontSubset: true FirstChar: 2 LastChar: 122 FontDescriptor: FontName: KRBWEG+URWPalladioL-Ital Flags: Symbolic FontBBox: -170, -305, 1010, 941 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: WMLZPF+URWPalladioL-Bold FontSubset: true FirstChar: 3 LastChar: 241 FontDescriptor: FontName: WMLZPF+URWPalladioL-Bold Flags: Symbolic FontBBox: -152, -301, 1000, 935 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: ILUGHX+VnURWPalladioL-Bold FontSubset: true FirstChar: 129 LastChar: 189 FontDescriptor: 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