| Literature DB >> 33686301 |
Wesley B Asher1,2,3, Peter Geggier1,3, Michael D Holsey3,4, Grant T Gilmore5, Avik K Pati6, Jozsef Meszaros1,3, Daniel S Terry6, Signe Mathiasen1,3, Megan J Kaliszewski5, Mitchell D McCauley5, Alekhya Govindaraju3, Zhou Zhou7,8, Kaleeckal G Harikumar9, Khuloud Jaqaman10,11, Laurence J Miller9, Adam W Smith5, Scott C Blanchard12,13, Jonathan A Javitch14,15,16,17.
Abstract
Class C G protein-coupled receptors (GPCRs) are known to form stable homodimers or heterodimers critical for function, but the oligomeric status of class A and B receptors, which constitute >90% of all GPCRs, remains hotly debated. Single-molecule fluorescence resonance energy transfer (smFRET) is a powerful approach with the potential to reveal valuable insights into GPCR organization but has rarely been used in living cells to study protein systems. Here, we report generally applicable methods for using smFRET to detect and track transmembrane proteins diffusing within the plasma membrane of mammalian cells. We leverage this in-cell smFRET approach to show agonist-induced structural dynamics within individual metabotropic glutamate receptor dimers. We apply these methods to representative class A, B and C receptors, finding evidence for receptor monomers, density-dependent dimers and constitutive dimers, respectively.Entities:
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Year: 2021 PMID： 33686301 PMCID： PMC8232828 DOI： 10.1038/s41592-021-01081-y
Source DB: PubMed Journal: Nat Methods ISSN： 1548-7091 Impact factor: 28.547