Publication details
Structural Basis for the 14-3-3 Protein-Dependent Inhibition of Phosducin Function
| Authors | |
|---|---|
| Year of publication | 2017 |
| Type | Article in Periodical |
| Magazine / Source | Biophysical Journal |
| MU Faculty or unit | |
| Citation | |
| Web | https://www.sciencedirect.com/science/article/pii/S0006349517302515?via%3Dihub |
| Doi | http://dx.doi.org/10.1016/j.bpj.2017.02.036 |
| Keywords | EXCHANGE-MASS-SPECTROMETRY; X-RAY-SCATTERING; INTRINSICALLY DISORDERED PROTEINS; SMALL-ANGLE SCATTERING; TRANSDUCIN BETA-GAMMA; LIGAND-BINDING; BIOLOGICAL MACROMOLECULES; PHOSPHORYLATION; REGULATOR; COMPLEX |
| Description | Phosducin (Pdc) is a conserved phosphoprotein that, when unphosphorylated, binds with high affinity to the complex of bg-subunits of G protein transducin (G(t beta gamma)). The ability of Pdc to bind to G(t beta gamma) is inhibited through its phosphorylation at S54 andS73 within the N-terminaldomain (Pdc-ND) followed by association with the scaffolding protein 14-3-3. However, the molecular basis for the 14-3-3-dependent inhibition of Pdc binding to G(t beta gamma) is unclear. By using small-angle x-ray scattering, high-resolution NMR spectroscopy, and limited proteolysis coupled with mass spectrometry, we show that phosphorylated Pdc and 14-3-3 forma complex in which the Pdc-ND region 45-80, which forms a part of Pdc's G(t beta gamma) binding surface and contains both phosphorylation sites, is restrained within the central channel of the 14-3-3 dimer, with both 14-3-3 binding motifs simultaneously participating in protein association. The N-terminal part of Pdc-NDis likely located outside the central channel of the 14-3-3 dimer, but Pdc residues 20-30, which are also involved in G(t beta gamma) binding, are positioned close to the surface of the 14-3-3 dimer. The C-terminal domain of Pdc is located outside the central channel and its structure is unaffected by the complex formation. These results indicate that the 14-3-3 protein-mediated inhibition of Pdc binding to Gtbg is based on steric occlusion of Pdc's G(t beta gamma) binding surface. |
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