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Journal: Science Advances
Article Title: Quantifying 3D live-cell membrane dynamics using dynamic metal-induced energy transfer spectroscopy (dynaMIET)
doi: 10.1126/sciadv.aed9613
Figure Lengend Snippet: ( A ) Schematic of the dynaMIET experiment conducted on a PM-labeled cell on top of a MIET substrate. ( B and E ) Same as , but for the PM system. n = 40 independent measurements over three independent experiments for living cells, and n = 25 independent measurements over two independent experiments for fixed cells. Data are obtained from independent measurements on multiple cells. For each cell, only one to two independent measurements were collected to avoid pseudo-replication. Independent experiments refer to measurements performed on different days using independently prepared samples. ( C and F ) Box plots of mean height values h 0 , diffusion coefficients D , fluctuation amplitudes ψ , and relaxation times τ ∗ for living and fixed cells. ( D ) Reconstructed 3D topography of the fluorescently labeled basal PM of a living COS-7 cell on a MIET substrate. The bottom panel offers an alternative viewing angle. Height is color-coded for clarity.
Article Snippet:
Techniques: Labeling, Diffusion-based Assay
Journal: Science Advances
Article Title: Quantifying 3D live-cell membrane dynamics using dynamic metal-induced energy transfer spectroscopy (dynaMIET)
doi: 10.1126/sciadv.aed9613
Figure Lengend Snippet: ( A , B , C , E , and F ) Same as but for the ER system. n = 25 independent measurements over 3 independent experiments for both living cells and fixed cells. Experimental replication and statistical details are the same as described in . ( D ) Exemplary 3D reconstructions (height maps) of fluorescently labeled ER in a live COS-7 cell on a MIET substrate. The xy image shows the fluorescence intensity. The bottom panel shows the height image from a different viewing angle.
Article Snippet:
Techniques: Labeling, Fluorescence
Journal: Science Advances
Article Title: Quantifying 3D live-cell membrane dynamics using dynamic metal-induced energy transfer spectroscopy (dynaMIET)
doi: 10.1126/sciadv.aed9613
Figure Lengend Snippet: ( A to F ) Same as but for the NE system. n = 33 independent measurements over three independent experiments. Experimental replication and statistical details are the same as described in . (B) iACFs obtained from living eGFP-nup153–modified COS-7 cell. The green solid line represents a fit with . The inset shows measured g i for a fixed cell. (E) Calculated hACF for a living and a fixed cell (inset).
Article Snippet:
Techniques: Modification
Journal: bioRxiv
Article Title: Pharmacological proximities in the GPCR family discovered using contact-informed amino-acid and binding pocket similarities
doi: 10.64898/2026.05.02.720972
Figure Lengend Snippet: a , The pharmacological neighborhood of the β 2 adrenoceptor. The transmembrane sequence similarity of 285 Class A GPCRs to the β 2 adrenoceptor are plotted against their corresponding GPCR-CoINPocket v2 scores. GPCRs are coloured broadly by classification: aminergics (red), peptide (purple), orphans (gray), and others (navy). b , Chemical structures of micromolar affinity ligands assessed in this study. Observed K i (affinity) of experimental ligands (green) or propranolol (blue) from competitive radioligand binding are displayed above each structure along with the PSICHIC-predicted K i . c , Competitive radioligand binding assays using putative surrogate ligands. Binding assays used crude purified membranes of COS-1 cells transiently transfected with β 2 adrenoceptor. Unlabelled competitor ligands were titrated against 0.5 nM [ 3 H]DHA and their affinity derived. Points and bars represent means ± SEM of n=3 experiments performed in triplicate. Curves were fit to a logistic single site binding model using GraphPad Prism v9. d , Boltz-1 and AlphaFold 3-predicted binding poses of propranolol and the low micromolar affinity off-target ligands co-folded with the β 2 adrenoceptor and their cognate receptors. In blue, the binding pose of the β 2 antagonist propranolol is shown bound to the β 2 adrenoceptor.
Article Snippet:
Techniques: Sequencing, Binding Assay, Purification, Transfection, Derivative Assay
Journal: bioRxiv
Article Title: Pharmacological proximities in the GPCR family discovered using contact-informed amino-acid and binding pocket similarities
doi: 10.64898/2026.05.02.720972
Figure Lengend Snippet: a , Competitive radioligand binding assay using putative surrogate ligands. Binding assays used crude purified membranes of COS-1 cells transiently transfected with β 2 adrenoceptor. Unlabelled competitor ligands were titrated against 0.5 nM [ 3 H]DHA and their affinity derived. Points and bars represent means ± SEMs of n=3 experiments performed in triplicate. Curves were fit to a logistic single site binding model using GraphPad Prism v9. b , Chemical structures of weakly-binding or inactive compounds assessed in this study. Observed K i (affinity) of experimental ligands from competitive radioligand binding > 1 μM (purple) or propranolol (blue) are displayed above each structure along with their PSICHIC-predicted K i . c , Radioligand dissociation assay of hit compounds in comparison to bona fide orthosteric antagonist propranolol. The rate at which [ 3 H]DHA dissociated from the β 2 adrenoceptor was measured over the course of 2 hrs at room temperature upon addition of 10 μM propranolol (blue) in the presence or absence of off-target compounds: 6 μM ATC 0175 (green) or 30 μM WAY 267464 (purple). Points and bars represent means ± SEMs of n=2-4 experiments performed in triplicate. Curves were fit to a logistic single site binding model using GraphPad Prism v9.
Article Snippet:
Techniques: Radio Ligand Binding Assay, Binding Assay, Purification, Transfection, Derivative Assay, Comparison