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

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F67985823%3A_____%2F17%3A00475511" target="_blank" >RIV/67985823:_____/17:00475511 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/61388971:_____/17:00475511 RIV/00216208:11310/17:10364663 RIV/00216224:14740/17:00100398

Výsledek na webu

<a href="http://dx.doi.org/10.1016/j.bpj.2017.02.036" target="_blank" >http://dx.doi.org/10.1016/j.bpj.2017.02.036</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.bpj.2017.02.036" target="_blank" >10.1016/j.bpj.2017.02.036</a>

Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

Phosducin (Pdc) is a conserved phosphoprotein that, when unphosphorylated, binds with high affinity to the complex of beta gamma subunits of G protein transducin. The ability of Pdc to bind to Gt(beta gamma) is inhibited through its phosphorylation at S54 andS73 within the N-terminal domain (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 Gt(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 Gt(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 Gt(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 Gt(beta gamma) is based on steric occlusion of Pdc's Gt(beta gamma) binding surface.

Název v anglickém jazyce

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

Popis výsledku anglicky

Phosducin (Pdc) is a conserved phosphoprotein that, when unphosphorylated, binds with high affinity to the complex of beta gamma subunits of G protein transducin. The ability of Pdc to bind to Gt(beta gamma) is inhibited through its phosphorylation at S54 andS73 within the N-terminal domain (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 Gt(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 Gt(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 Gt(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 Gt(beta gamma) is based on steric occlusion of Pdc's Gt(beta gamma) binding surface.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10609 - Biochemical research methods

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2017

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Biophysical Journal

ISSN

0006-3495

e-ISSN

—

Svazek periodika

112

Číslo periodika v rámci svazku

7

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

11

Strana od-do

1339-1349

Kód UT WoS článku

000398956000007

EID výsledku v databázi Scopus

2-s2.0-85017311104