Figure 6: 19F-NMR spectra revealed the structural alteration of β-arrestin-1 in response to the binding of Clathrin. | Nature Communications

Figure 6: 19F-NMR spectra revealed the structural alteration of β-arrestin-1 in response to the binding of Clathrin.

From: Phospho-selective mechanisms of arrestin conformations and functions revealed by unnatural amino acid incorporation and 19F-NMR

Figure 6

Chemical shifts after phosphopeptide/β-arrestin-1/clathrin ternary complex formation at specific F2Y incorporation sites: at N375-F2Y close to the classic clathrin-binding box (a), at L388 in the splice loop (b) and at Y249-F2Y (c). (d) No significant changes (Δp.p.m.<0.05) were observed for the chemical shifts at F277-F2Y position in the 19F-NMR spectra after clathrin binding. (e) Paramagnetic titration experiments. As the concentration of the Cr increased, the full width at half maximum of the 1D-19F-NMR spectra at the F2Y-N375 position of the β-arrestin-1/GRK2Bpp complex significantly increased (Δp.p.m.=25 HZ), followed by the β-arrestin-1/GRK6 complex in the presence of clathrin (Δp.p.m.=16 HZ), and then the β-arrestin-1/GRK2Bpp/clathrin ternary complex (Δp.p.m.=5 HZ) and the β-arrestin-1 alone (Δp.p.m.=4 HZ), indicating a greater protective effect of Cr at the N375 position of the β-arrestin-1/GRK2Bpp complex in the absence of clathrin. (f) A cartoon illustration of the phospho-pattern-encoded structural rearrangements of β-arrestin-1 and its subsequent stabilization by clathrin binding. The red ball indicates the phosphate. Although the binding of GRK2App, GRK2Bpp and V2Rpp all caused a partial dislodging of the C terminus of β-arrestin-1, the resulting structural states induced by different phosphopeptides could be different. Clathrin binding stabilized all these phosphopeptide-occupied arrestins to a single state.

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