About: Identification of a Preassembled TRH Receptor-G(q/11) Protein Complex in HEK293 Cells     Goto   Sponge   NotDistinct   Permalink

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  • Protein-protein interactions define specificity in signal transduction and these interactions are central to transmembrane signaling by G-protein-coupled receptors (GPCRs). It is not quite clear, however, whether GPCRs and the regulatory trimeric G-proteins behave as freely and independently diffusible molecules in the plasma membrane or whether they form some preassociated complexes. Here we used clear-native polyacrylamide gel electrophoresis (CN-PAGE) to investigate the presumed coupling between thyrotropin-releasing hormone (TRH) receptor and its cognate G(q/11) protein in HEK293 cells expressing high levels of these proteins. Under different solubilization conditions, the TRH receptor (TRH-R) was identified to form a putative pentameric complex composed of TRH-R homodimer and G(q/11) protein. The presumed association of TRH-R with G(q/11)alpha or G beta proteins in plasma membranes was verified by RNAi experiments. After 10- or 30-min hormone treatment, TRH-R signaling complexes gradually dissociated with a concomitant release of receptor homodimers. These observations support the model in which GPCRs can be coupled to trimeric G-proteins in preassembled signaling complexes, which might be dynamically regulated upon receptor activation. The precoupling of receptors with their cognate G-proteins can contribute to faster G-protein activation and subsequent signal transfer into the cell interior.
  • Protein-protein interactions define specificity in signal transduction and these interactions are central to transmembrane signaling by G-protein-coupled receptors (GPCRs). It is not quite clear, however, whether GPCRs and the regulatory trimeric G-proteins behave as freely and independently diffusible molecules in the plasma membrane or whether they form some preassociated complexes. Here we used clear-native polyacrylamide gel electrophoresis (CN-PAGE) to investigate the presumed coupling between thyrotropin-releasing hormone (TRH) receptor and its cognate G(q/11) protein in HEK293 cells expressing high levels of these proteins. Under different solubilization conditions, the TRH receptor (TRH-R) was identified to form a putative pentameric complex composed of TRH-R homodimer and G(q/11) protein. The presumed association of TRH-R with G(q/11)alpha or G beta proteins in plasma membranes was verified by RNAi experiments. After 10- or 30-min hormone treatment, TRH-R signaling complexes gradually dissociated with a concomitant release of receptor homodimers. These observations support the model in which GPCRs can be coupled to trimeric G-proteins in preassembled signaling complexes, which might be dynamically regulated upon receptor activation. The precoupling of receptors with their cognate G-proteins can contribute to faster G-protein activation and subsequent signal transfer into the cell interior. (en)
Title
  • Identification of a Preassembled TRH Receptor-G(q/11) Protein Complex in HEK293 Cells
  • Identification of a Preassembled TRH Receptor-G(q/11) Protein Complex in HEK293 Cells (en)
skos:prefLabel
  • Identification of a Preassembled TRH Receptor-G(q/11) Protein Complex in HEK293 Cells
  • Identification of a Preassembled TRH Receptor-G(q/11) Protein Complex in HEK293 Cells (en)
skos:notation
  • RIV/00216208:11310/12:10123864!RIV13-GA0-11310___
http://linked.open...avai/predkladatel
http://linked.open...avai/riv/aktivita
http://linked.open...avai/riv/aktivity
  • P(GD305/08/H037), S, Z(MSM0021620858)
http://linked.open...iv/cisloPeriodika
  • 37
http://linked.open...vai/riv/dodaniDat
http://linked.open...aciTvurceVysledku
http://linked.open.../riv/druhVysledku
http://linked.open...iv/duvernostUdaju
http://linked.open...titaPredkladatele
http://linked.open...dnocenehoVysledku
  • 140276
http://linked.open...ai/riv/idVysledku
  • RIV/00216208:11310/12:10123864
http://linked.open...riv/jazykVysledku
http://linked.open.../riv/klicovaSlova
  • protein complex; clear-native electrophoresis; g(q/11) protein; Thyrotropin-releasing hormone receptor (en)
http://linked.open.../riv/klicoveSlovo
http://linked.open...odStatuVydavatele
  • JP - Japonsko
http://linked.open...ontrolniKodProRIV
  • [8C81FFC261B9]
http://linked.open...i/riv/nazevZdroje
  • Cell Structure and Function
http://linked.open...in/vavai/riv/obor
http://linked.open...ichTvurcuVysledku
http://linked.open...cetTvurcuVysledku
http://linked.open...vavai/riv/projekt
http://linked.open...UplatneniVysledku
http://linked.open...v/svazekPeriodika
  • 37
http://linked.open...iv/tvurceVysledku
  • Novotný, Jiří
  • Drastichová, Zdeňka
http://linked.open...ain/vavai/riv/wos
  • 000299108700001
http://linked.open...n/vavai/riv/zamer
issn
  • 0386-7196
number of pages
http://localhost/t...ganizacniJednotka
  • 11310
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