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

J 2017

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

KACIROVA, M., Jiří NOVÁČEK, P. MAN, V. OBSILOVA, T. OBSIL et. al.

Základní údaje

Originální název

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

Autoři

KACIROVA, M. (203 Česká republika), Jiří NOVÁČEK (203 Česká republika, garant, domácí), P. MAN (203 Česká republika), V. OBSILOVA (203 Česká republika) a T. OBSIL (203 Česká republika)

Vydání

Biophysical Journal, Bethesda, USA, Biophysical Society, 2017, 0006-3495

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10610 Biophysics

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 3.495

Kód RIV

RIV/00216224:14740/17:00100398

Organizační jednotka

Středoevropský technologický institut

DOI

http://dx.doi.org/10.1016/j.bpj.2017.02.036

UT WoS

000398956000007

Klíčová slova anglicky

EXCHANGE-MASS-SPECTROMETRY; X-RAY-SCATTERING; INTRINSICALLY DISORDERED PROTEINS; SMALL-ANGLE SCATTERING; TRANSDUCIN BETA-GAMMA; LIGAND-BINDING; BIOLOGICAL MACROMOLECULES; PHOSPHORYLATION; REGULATOR; COMPLEX

Štítky

CF NMR, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 16. 3. 2018 16:24, Mgr. Pavla Foltynová, Ph.D.

Anotace

V originále

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.

Návaznosti

LM2015043, projekt VaV

Název: Česká infrastruktura pro integrativní strukturní biologii (Akronym: CIISB)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Czech Infrastructure for Integrative Structural Biology

Citovat

KACIROVA, M., Jiří NOVÁČEK, P. MAN, V. OBSILOVA a T. OBSIL. Structural Basis for the 14-3-3 Protein-Dependent Inhibition of Phosducin Function. Biophysical Journal. Bethesda, USA: Biophysical Society, 2017, roč. 112, č. 7, s. 1339-1349. ISSN 0006-3495. Dostupné z: https://dx.doi.org/10.1016/j.bpj.2017.02.036.

@article{1411420,
   author = {Kacirova, M. and Nováček, Jiří and Man, P. and Obsilova, V. and Obsil, T.},
   article_location = {Bethesda, USA},
   article_number = {7},
   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},
   language = {eng},
   issn = {0006-3495},
   journal = {Biophysical Journal},
   title = {Structural Basis for the 14-3-3 Protein-Dependent Inhibition of Phosducin Function},
   url = {https://www.sciencedirect.com/science/article/pii/S0006349517302515?via%3Dihub},
   volume = {112},
   year = {2017}
}

TY  - JOUR
ID  - 1411420
AU  - Kacirova, M. - Nováček, Jiří - Man, P. - Obsilova, V. - Obsil, T.
PY  - 2017
TI  - Structural Basis for the 14-3-3 Protein-Dependent Inhibition of Phosducin Function
JF  - Biophysical Journal
VL  - 112
IS  - 7
SP  - 1339-1349
EP  - 1339-1349
PB  - Biophysical Society
SN  - 00063495
KW  - EXCHANGE-MASS-SPECTROMETRY
KW  - X-RAY-SCATTERING
KW  - INTRINSICALLY DISORDERED PROTEINS
KW  - SMALL-ANGLE SCATTERING
KW  - TRANSDUCIN BETA-GAMMA
KW  - LIGAND-BINDING
KW  - BIOLOGICAL MACROMOLECULES
KW  - PHOSPHORYLATION
KW  - REGULATOR
KW  - COMPLEX
UR  - https://www.sciencedirect.com/science/article/pii/S0006349517302515?via%3Dihub
N2  - 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.
ER  -

KACIROVA, M., Jiří NOVÁČEK, P. MAN, V. OBSILOVA a T. OBSIL. Structural Basis for the 14-3-3 Protein-Dependent Inhibition of Phosducin Function. \textit{Biophysical Journal}. Bethesda, USA: Biophysical Society, 2017, roč.~112, č.~7, s.~1339-1349. ISSN~0006-3495. Dostupné z: https://dx.doi.org/10.1016/j.bpj.2017.02.036.

Podrobný výpis o publikaci