Identification of a preassembled TRH receptor-G(q/11) protein complex in HEK293 cells
Jazyk angličtina Země Japonsko Médium print
Typ dokumentu časopisecké články, práce podpořená grantem
PubMed
22240728
DOI
10.1247/csf.11024
PII: JST.JSTAGE/csf/11024
Knihovny.cz E-zdroje
- MeSH
- buněčná membrána metabolismus MeSH
- buněčné linie MeSH
- HEK293 buňky MeSH
- hormon uvolňující thyreotropin genetika metabolismus MeSH
- lidé MeSH
- proteiny vázající GTP - alfa-podjednotky Gq-G11 chemie genetika metabolismus MeSH
- receptory thyroliberinu chemie genetika metabolismus MeSH
- signální transdukce MeSH
- transfekce MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- hormon uvolňující thyreotropin MeSH
- proteiny vázající GTP - alfa-podjednotky Gq-G11 MeSH
- receptory thyroliberinu MeSH
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)α or Gβ 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.
Citace poskytuje Crossref.org
Biochemical and physiological insights into TRH receptor-mediated signaling