期刊
NATURE METHODS
卷 18, 期 4, 页码 397-+出版社
NATURE RESEARCH
DOI: 10.1038/s41592-021-01081-y
关键词
-
资金
- NIH [7R01GM098859-09, MH54137, R15EY024451, R35GM119619]
- Hope for Depression Research Foundation
- Lieber Center for Schizophrenia Research
- Brain and Behavior Research Foundation NARSAD Young Investigator Award
- National Science Foundation [CHE-1753060]
- UTSW Endowed Scholars Program
- Single-Molecule Imaging Center at St. Jude Children's Research Hospital
This study presents a generally applicable method for using single-molecule fluorescence resonance energy transfer (smFRET) to detect and track transmembrane proteins diffusing within the plasma membrane of mammalian cells. The in-cell smFRET approach reveals agonist-induced structural dynamics within individual metabotropic glutamate receptor dimers. Evidence for receptor monomers, density-dependent dimers, and constitutive dimers in representative class A, B, and C receptors were observed using these methods.
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.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据