期刊
SINGLE MOLECULE SPECTROSCOPY AND SUPERRESOLUTION IMAGING XII
卷 10884, 期 -, 页码 -出版社
SPIE-INT SOC OPTICAL ENGINEERING
DOI: 10.1117/12.2507546
关键词
Neurotensin receptor 1; single molecule spectroscopy; SMALP; ABELtrap; homoFRET; anisotropy
资金
- State of Thuringia [FKZ 12026-515]
- Jena University Hospital
- Deutsche Forschungsgemeinschaft DFG in the Collaborative Research Center [Transregio 166]
- DFG [BO1891/10-2, BO1891/15-1, BO1891/16-1, BO1891/18-2]
- ACP Explore project within the ProExcellence initiative ACP2020 from the State of Thuringia
G protein-coupled receptors (GPCRs) are a large superfamily of membrane proteins that are activated by extracellular small molecules or photons. Neurotensin receptor 1 (NTSR1) is a GPCR that is activated by neurotensin, i.e. a 13 amino acid peptide. Binding of neurotensin induces conformational changes in the receptor that trigger the intracellular signaling processes. While recent single-molecule studies have reported a dynamic monomer -dimer equilibrium of NTSR1 in vitro, a biophysical characterization of the oligomerization status of NTSR1 in living mammalian cells is complicated. Here we report on the oligomerization state of the human NTSR1 tagged with mRuby3 by dissolving the plasma membranes of living HEK293T cells into 10 nm-sized soluble lipid nanoparticles by addition of styrene-maleic acid copolymers (SMALPs). Single SMALPs were analyzed one after another in solution by multi-parameter single molecule spectroscopy including brightness, fluorescence lifetime and anisotropy for homoFRET. Brightness analysis was improved using single SMALP detection in a confocal ABELtrap for extended observation times in solution. A bimodal brightness distribution indicated a significant fraction of dimeric NTSR1 in SMALPs or in the plasma membrane, respectively, before addition of neurotensin.
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