Structural Basis for the 14-3-3 Protein-Dependent Inhibition of Phosducin Function

KACIROVA, M., Jiří NOVÁČEK, P. MAN, V. OBSILOVA and T. OBSIL. Structural Basis for the 14-3-3 Protein-Dependent Inhibition of Phosducin Function. Biophysical Journal. Bethesda, USA: Biophysical Society, 2017, vol. 112, No 7, p. 1339-1349. ISSN 0006-3495. Available from: https://dx.doi.org/10.1016/j.bpj.2017.02.036.

Basic information

• Original name: Structural Basis for the 14-3-3 Protein-Dependent Inhibition of Phosducin Function
• Authors: KACIROVA, M. (203 Czech Republic), Jiří NOVÁČEK (203 Czech Republic, guarantor, belonging to the institution), P. MAN (203 Czech Republic), V. OBSILOVA (203 Czech Republic) and T. OBSIL (203 Czech Republic).
• Edition: Biophysical Journal, Bethesda, USA, Biophysical Society, 2017, 0006-3495.

Other information

• Original language: English
• Type of outcome: Article in a journal
• Field of Study: 10610 Biophysics
• Country of publisher: United States of America
• Confidentiality degree: is not subject to a state or trade secret
• WWW: URL
• Impact factor: Impact factor: 3.495
• RIV identification code: RIV/00216224:14740/17:00100398
• Organization unit: Central European Institute of Technology
• Doi: http://dx.doi.org/10.1016/j.bpj.2017.02.036
• UT WoS: 000398956000007
• Keywords in English: EXCHANGE-MASS-SPECTROMETRY; X-RAY-SCATTERING; INTRINSICALLY DISORDERED PROTEINS; SMALL-ANGLE SCATTERING; TRANSDUCIN BETA-GAMMA; LIGAND-BINDING; BIOLOGICAL MACROMOLECULES; PHOSPHORYLATION; REGULATOR; COMPLEX
• Tags: CF NMR, rivok
• Tags: International impact, Reviewed

Abstract

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.

Links

• LM2015043, research and development project: Name: Česká infrastruktura pro integrativní strukturní biologii (Acronym: CIISB)

Investor: Ministry of Education, Youth and Sports of the CR