• Je něco špatně v tomto záznamu ?

14-3-3 protein directly interacts with the kinase domain of calcium/calmodulin-dependent protein kinase kinase (CaMKK2)

K. Psenakova, O. Petrvalska, S. Kylarova, D. Lentini Santo, D. Kalabova, P. Herman, V. Obsilova, T. Obsil,

. 2018 ; 1862 (7) : 1612-1625. [pub] 20180410

Jazyk angličtina Země Nizozemsko

Typ dokumentu časopisecké články, práce podpořená grantem

Perzistentní odkaz   https://www.medvik.cz/link/bmc18033166

BACKGROUND: Calcium/calmodulin-dependent protein kinase kinase 2 (CaMKK2) is a member of the Ca2+/calmodulin-dependent kinase (CaMK) family involved in adiposity regulation, glucose homeostasis and cancer. This upstream activator of CaMKI, CaMKIV and AMP-activated protein kinase is inhibited by phosphorylation, which also triggers an association with the scaffolding protein 14-3-3. However, the role of 14-3-3 in the regulation of CaMKK2 remains unknown. METHODS: The interaction between phosphorylated CaMKK2 and the 14-3-3γ protein, as well as the architecture of their complex, were studied using enzyme activity measurements, small-angle x-ray scattering (SAXS), time-resolved fluorescence spectroscopy and protein crystallography. RESULTS: Our data suggest that the 14-3-3 protein binding does not inhibit the catalytic activity of phosphorylated CaMKK2 but rather slows down its dephosphorylation. Structural analysis indicated that the complex is flexible and that CaMKK2 is located outside the phosphopeptide-binding central channel of the 14-3-3γ dimer. Furthermore, 14-3-3γ appears to interact with and affect the structure of several regions of CaMKK2 outside the 14-3-3 binding motifs. In addition, the structural basis of interactions between 14-3-3 and the 14-3-3 binding motifs of CaMKK2 were elucidated by determining the crystal structures of phosphopeptides containing these motifs bound to 14-3-3. CONCLUSIONS: 14-3-3γ protein directly interacts with the kinase domain of CaMKK2 and the region containing the inhibitory phosphorylation site Thr145 within the N-terminal extension. GENERAL SIGNIFICANCE: Our results suggested that CaMKK isoforms differ in their 14-3-3-mediated regulations and that the interaction between 14-3-3 protein and the N-terminal 14-3-3-binding motif of CaMKK2 might be stabilized by small-molecule compounds.

Citace poskytuje Crossref.org

000      
00000naa a2200000 a 4500
001      
bmc18033166
003      
CZ-PrNML
005      
20181017152535.0
007      
ta
008      
181008s2018 ne f 000 0|eng||
009      
AR
024    7_
$a 10.1016/j.bbagen.2018.04.006 $2 doi
035    __
$a (PubMed)29649512
040    __
$a ABA008 $b cze $d ABA008 $e AACR2
041    0_
$a eng
044    __
$a ne
100    1_
$a Psenakova, Katarina $u Department of Physical and Macromolecular Chemistry, Faculty of Science, Charles University, Prague, Czech Republic; BioCeV - Institute of Physiology, The Czech Academy of Sciences, Vestec, Czech Republic.
245    10
$a 14-3-3 protein directly interacts with the kinase domain of calcium/calmodulin-dependent protein kinase kinase (CaMKK2) / $c K. Psenakova, O. Petrvalska, S. Kylarova, D. Lentini Santo, D. Kalabova, P. Herman, V. Obsilova, T. Obsil,
520    9_
$a BACKGROUND: Calcium/calmodulin-dependent protein kinase kinase 2 (CaMKK2) is a member of the Ca2+/calmodulin-dependent kinase (CaMK) family involved in adiposity regulation, glucose homeostasis and cancer. This upstream activator of CaMKI, CaMKIV and AMP-activated protein kinase is inhibited by phosphorylation, which also triggers an association with the scaffolding protein 14-3-3. However, the role of 14-3-3 in the regulation of CaMKK2 remains unknown. METHODS: The interaction between phosphorylated CaMKK2 and the 14-3-3γ protein, as well as the architecture of their complex, were studied using enzyme activity measurements, small-angle x-ray scattering (SAXS), time-resolved fluorescence spectroscopy and protein crystallography. RESULTS: Our data suggest that the 14-3-3 protein binding does not inhibit the catalytic activity of phosphorylated CaMKK2 but rather slows down its dephosphorylation. Structural analysis indicated that the complex is flexible and that CaMKK2 is located outside the phosphopeptide-binding central channel of the 14-3-3γ dimer. Furthermore, 14-3-3γ appears to interact with and affect the structure of several regions of CaMKK2 outside the 14-3-3 binding motifs. In addition, the structural basis of interactions between 14-3-3 and the 14-3-3 binding motifs of CaMKK2 were elucidated by determining the crystal structures of phosphopeptides containing these motifs bound to 14-3-3. CONCLUSIONS: 14-3-3γ protein directly interacts with the kinase domain of CaMKK2 and the region containing the inhibitory phosphorylation site Thr145 within the N-terminal extension. GENERAL SIGNIFICANCE: Our results suggested that CaMKK isoforms differ in their 14-3-3-mediated regulations and that the interaction between 14-3-3 protein and the N-terminal 14-3-3-binding motif of CaMKK2 might be stabilized by small-molecule compounds.
650    _2
$a proteiny 14-3-3 $x metabolismus $7 D048948
650    _2
$a proteinkinasy aktivované AMP $x metabolismus $7 D055372
650    _2
$a aminokyselinové motivy $7 D020816
650    _2
$a kinasa proteinkinasy závislé na vápníku a kalmodulinu $x metabolismus $7 D054737
650    _2
$a proteinkinasa závislá na vápníku a kalmodulinu typ 1 $x metabolismus $7 D054729
650    _2
$a lidé $7 D006801
650    _2
$a molekulární modely $7 D008958
650    _2
$a fosforylace $x účinky léků $7 D010766
650    _2
$a vazba proteinů $7 D011485
650    _2
$a konformace proteinů $x účinky léků $7 D011487
650    _2
$a proteinové domény $7 D000072417
650    _2
$a mapování interakce mezi proteiny $7 D025941
650    _2
$a posttranslační úpravy proteinů $7 D011499
650    _2
$a rekombinantní proteiny $x metabolismus $7 D011994
655    _2
$a časopisecké články $7 D016428
655    _2
$a práce podpořená grantem $7 D013485
700    1_
$a Petrvalska, Olivia $u Department of Physical and Macromolecular Chemistry, Faculty of Science, Charles University, Prague, Czech Republic; BioCeV - Institute of Physiology, The Czech Academy of Sciences, Vestec, Czech Republic.
700    1_
$a Kylarova, Salome $u Department of Physical and Macromolecular Chemistry, Faculty of Science, Charles University, Prague, Czech Republic; BioCeV - Institute of Physiology, The Czech Academy of Sciences, Vestec, Czech Republic.
700    1_
$a Lentini Santo, Domenico $u Department of Physical and Macromolecular Chemistry, Faculty of Science, Charles University, Prague, Czech Republic.
700    1_
$a Kalabova, Dana $u BioCeV - Institute of Physiology, The Czech Academy of Sciences, Vestec, Czech Republic.
700    1_
$a Herman, Petr $u Institute of Physics, Faculty of Mathematics and Physics, Charles University, Prague, Czech Republic.
700    1_
$a Obsilova, Veronika $u BioCeV - Institute of Physiology, The Czech Academy of Sciences, Vestec, Czech Republic. Electronic address: veronika.obsilova@fgu.cas.cz.
700    1_
$a Obsil, Tomas $u Department of Physical and Macromolecular Chemistry, Faculty of Science, Charles University, Prague, Czech Republic; BioCeV - Institute of Physiology, The Czech Academy of Sciences, Vestec, Czech Republic. Electronic address: obsil@natur.cuni.cz.
773    0_
$w MED00000717 $t Biochimica et biophysica acta. General subjects $x 0304-4165 $g Roč. 1862, č. 7 (2018), s. 1612-1625
856    41
$u https://pubmed.ncbi.nlm.nih.gov/29649512 $y Pubmed
910    __
$a ABA008 $b sig $c sign $y a $z 0
990    __
$a 20181008 $b ABA008
991    __
$a 20181017153034 $b ABA008
999    __
$a ok $b bmc $g 1339384 $s 1030160
BAS    __
$a 3
BAS    __
$a PreBMC
BMC    __
$a 2018 $b 1862 $c 7 $d 1612-1625 $e 20180410 $i 0304-4165 $m Biochimica et biophysica acta. G, General subjects $n Biochem Biophys Acta $x MED00000717
LZP    __
$a Pubmed-20181008

Najít záznam

Citační ukazatele

Pouze přihlášení uživatelé

Možnosti archivace