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Sequential inter- and intrasubunit rearrangements during activation of dimeric metabotropic glutamate receptor 1
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SYSNO ASEP 0381622 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Sequential inter- and intrasubunit rearrangements during activation of dimeric metabotropic glutamate receptor 1 Author(s) Hlaváčková, Veronika (UMG-J)
Zabel, U. (DE)
Franková, Daniela (UMG-J)
Batz, J. (DE)
Hoffmann, C. (DE)
Prezeau, L. (FR)
Pin, J. P. (FR)
Blahoš, Jaroslav (UMG-J) RID
Lohse, M. J. (DE)Source Title Science Signaling. - : American Association for the Advancement of Science - ISSN 1945-0877
Roč. 5, č. 237 (2012), ra59Number of pages 11 s. Publication form Online - E Language eng - English Country US - United States Keywords G-protein coupled receptor ; metabotropic glutamate receptor 1 ; class C GPCR Subject RIV EB - Genetics ; Molecular Biology R&D Projects GA303/08/1591 GA ČR - Czech Science Foundation (CSF) LC06063 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) GAP303/12/2408 GA ČR - Czech Science Foundation (CSF) CEZ AV0Z50520514 - UMG-J (2005-2011) UT WOS 000307885200002 DOI 10.1126/scisignal.2002720 Annotation The metabotropic glutamate receptor 1 (mGluR1), a class C member of the heterotrimeric guanine nucleotide-binding protein (G protein)-coupled receptor family, is a constitutive dimer that regulates excitatory neurotransmission. We investigated the role of homodimer formation in mGluR1 activation by examining activation-dependent inter-and intrasubunit conformational changes by fluorescence resonance energy transfer (FRET). We inserted yellow and cyan fluorescent proteins in the second intracellular loop and at the carboxyl terminus of mGluR1 to act as FRET sensors and expressed these proteins in human embryonic kidney 293 cells. Agonist-dependent activation of these mGluR1 chimeras rapidly increased the intersubunit FRET, suggesting rapid movement of the subunits relative to each other. After intersubunit movement, the intrasubunit FRET decreased, reflecting conformational changes within a subunit. Cotransfection of chimeric receptor subunits that were capable or incapable of G protein coupling revealed that only a single subunit assumes an active state in an mGluR1 receptor dimer. Workplace Institute of Molecular Genetics Contact Nikol Škňouřilová, nikol.sknourilova@img.cas.cz, Tel.: 241 063 217 Year of Publishing 2013
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