Fig. 1: Fluorescence spectroscopy resolves membrane receptor mobility. | Communications Biology

Fig. 1: Fluorescence spectroscopy resolves membrane receptor mobility.

From: Unraveling the hidden temporal range of fast β2-adrenergic receptor mobility by time-resolved fluorescence

Fig. 1

a CHO-K1 cells were grown on glass coverslips (top, fluorescent confocal maximum intensity projection vs. brightfield illumination) and transiently transfected with four different β2-AR constructs (middle) carrying three differently sized fluorophores (bottom): N-terminal EGFP or SNAPtag, EGFP at the intracellular loop 3 or Me-Tet-ATTO488 coupled to the unnatural amino acid trans-cycloctene (TCO) at position 186 (β2-ARA186TCO). b Structures of agonist (isoproterenol, ISO), partial agonist (salbutamol, SAL) or inverse agonist (carazolol, CAR) used in this study to compare β2-AR diffusion dynamics upon their addition to probe for possible changes in oligomerization or protein clustering. c The combination of three different time resolved spectroscopy experiments with overlapping temporal resolution allows to probe the translational and rotational dynamics in the range from ps to sec; Time-resolved anisotropy (TRA, top) is sensitive to rotation in the ps to the ns range, Fluorescence correlation spectroscopy (FCS, bottom) probes dynamics in the range from µs to s, while continuous-wave FCS (fullFCS, middle) covers a broad range from ps to s.

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