Fig. 3: βArr1 stabilizes pre-existing states in the presence of agonist and/or BAM. | Nature Communications

Fig. 3: βArr1 stabilizes pre-existing states in the presence of agonist and/or BAM.

From: Stabilization of pre-existing neurotensin receptor conformational states by β-arrestin-1 and the biased allosteric modulator ML314

Fig. 3: βArr1 stabilizes pre-existing states in the presence of agonist and/or BAM.

a Microscale thermophoresis (MST) measured the affinity of βArr1-3A for NT8-13:enNTS1ΔM4 as 0.99 ± 0.1 μM ( ± SEM) using a single-site quadratic binding model. Data was collected as n = 3 biologically independent experiments with 5 or 6 technical repeats. Source data are provided as a Source Data file. Overlays of (b) NT8-13:enNTS1ΔM4 (forest green) and NT8-13:enNTS1ΔM4:βArr1-3A (purple), (c) ML314:enNTS1ΔM4 (cyan) and ML314:enNTS1ΔM4:βArr1-3A (tan), and (d) NT8-13:ML314:enNTS1ΔM4 (maroon) and NT8-13:ML314:enNTS1ΔM4:βArr1-3A (royal blue) 1H-13C HMQC spectra. Transducer spectra included 2.3x molar equivalents βArr1-3A. Peaks marked with an asterisk represent natural abundance βArr1-3A M411 (also see Supplementary Fig. 2b). Extracted spectral region of (e) M2445.45 and (f) M2505.51 from NT8-13:enNTS1ΔM4:βArr1-3A (purple), ML314:enNTS1ΔM4:βArr1-3A (tan), and NT8-13:ML314:enNTS1ΔM4:βArr1-3A (royal blue) 1H-13C HMQC spectra. One dimensional 1H cross-sectional slices (corresponding to dotted line) shown on top. Dots denote the residue’s chemical shift position in spectra of the corresponding colour with additional dots shown for ligand-only spectra (NT8-13:enNTS1ΔM4, forest green; ML314:enNTS1ΔM4, cyan; NT8-13:ML314:enNTS1ΔM4, maroon). All spectra were recorded at 600 MHz with receptor concentrations of 66 μM.

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