Detail
Článek
Článek online
FT
Medvik - BMČ
  • Je něco špatně v tomto záznamu ?

Specific phosphorylation of microtubule-associated protein 2c by extracellular signal-regulated kinase reduces interactions at its Pro-rich regions

J. Plucarová, S. Jansen, S. Narasimhan, A. Laníková, M. Lewitzky, SM. Feller, L. Žídek

. 2022 ; 298 (10) : 102384. [pub] 20220817

Jazyk angličtina Země Spojené státy americké

Typ dokumentu časopisecké články, práce podpořená grantem

Perzistentní odkaz   https://www.medvik.cz/link/bmc22033180

Microtubule-associated protein 2 (MAP2) is an important neuronal target of extracellular signal-regulated kinase 2 (ERK2) involved in Raf signaling pathways, but mechanistic details of MAP2 phosphorylation are unclear. Here, we used NMR spectroscopy to quantitatively describe the kinetics of phosphorylation of individual serines and threonines in the embryonic MAP2 variant MAP2c. We carried out real-time monitoring of phosphorylation to discover major phosphorylation sites that were not identified in previous studies relying on specific antibodies. Our comparison with the phosphorylation of MAP2c by a model cyclin-dependent kinase CDK2 and with phosphorylation of the MAP2c homolog Tau revealed differences in phosphorylation profiles that explain specificity of regulation of biological functions of MAP2c and Tau. To probe the molecular basis of the regulatory effect of ERK2, we investigated the interactions of phosphorylated and unphosphorylated MAP2c by NMR with single-residue resolution. As ERK2 phosphorylates mostly outside the regions binding microtubules, we studied the binding of proteins other than tubulin, namely regulatory subunit RIIα of cAMP-dependent PKA, adapter protein Grb2, Src homology domain 3 of tyrosine kinases Fyn and Abl, and ERK2 itself. We found ERK2 phosphorylation interfered mostly with binding to proline-rich regions of MAP2c. Furthermore, our NMR experiments in SH-SY5Y neuroblastoma cell lysates showed that the kinetics of dephosphorylation are compatible with in-cell NMR studies and that residues targeted by ERK2 and PKA are efficiently phosphorylated in the cell lysates. Taken together, our results provide a deeper characterization of MAP2c phosphorylation and its effects on interactions with other proteins.

Citace poskytuje Crossref.org

000      
00000naa a2200000 a 4500
001      
bmc22033180
003      
CZ-PrNML
005      
20230131150641.0
007      
ta
008      
230120s2022 xxu f 000 0|eng||
009      
AR
024    7_
$a 10.1016/j.jbc.2022.102384 $2 doi
035    __
$a (PubMed)35987383
040    __
$a ABA008 $b cze $d ABA008 $e AACR2
041    0_
$a eng
044    __
$a xxu
100    1_
$a Plucarová, Jitka $u National Centre for Biomolecular Research, Faculty of Science, Masaryk University, Brno, Czech Republic; Central European Institute of Technology, Masaryk University, Brno, Czech Republic
245    10
$a Specific phosphorylation of microtubule-associated protein 2c by extracellular signal-regulated kinase reduces interactions at its Pro-rich regions / $c J. Plucarová, S. Jansen, S. Narasimhan, A. Laníková, M. Lewitzky, SM. Feller, L. Žídek
520    9_
$a Microtubule-associated protein 2 (MAP2) is an important neuronal target of extracellular signal-regulated kinase 2 (ERK2) involved in Raf signaling pathways, but mechanistic details of MAP2 phosphorylation are unclear. Here, we used NMR spectroscopy to quantitatively describe the kinetics of phosphorylation of individual serines and threonines in the embryonic MAP2 variant MAP2c. We carried out real-time monitoring of phosphorylation to discover major phosphorylation sites that were not identified in previous studies relying on specific antibodies. Our comparison with the phosphorylation of MAP2c by a model cyclin-dependent kinase CDK2 and with phosphorylation of the MAP2c homolog Tau revealed differences in phosphorylation profiles that explain specificity of regulation of biological functions of MAP2c and Tau. To probe the molecular basis of the regulatory effect of ERK2, we investigated the interactions of phosphorylated and unphosphorylated MAP2c by NMR with single-residue resolution. As ERK2 phosphorylates mostly outside the regions binding microtubules, we studied the binding of proteins other than tubulin, namely regulatory subunit RIIα of cAMP-dependent PKA, adapter protein Grb2, Src homology domain 3 of tyrosine kinases Fyn and Abl, and ERK2 itself. We found ERK2 phosphorylation interfered mostly with binding to proline-rich regions of MAP2c. Furthermore, our NMR experiments in SH-SY5Y neuroblastoma cell lysates showed that the kinetics of dephosphorylation are compatible with in-cell NMR studies and that residues targeted by ERK2 and PKA are efficiently phosphorylated in the cell lysates. Taken together, our results provide a deeper characterization of MAP2c phosphorylation and its effects on interactions with other proteins.
650    _2
$a lidé $7 D006801
650    12
$a extracelulárním signálem regulované MAP kinasy $x metabolismus $7 D048049
650    12
$a proteiny asociované s mikrotubuly $x metabolismus $7 D008869
650    _2
$a mikrotubuly $x metabolismus $7 D008870
650    _2
$a fosforylace $7 D010766
650    12
$a proteinkinasy řízené prolinem $x metabolismus $7 D038461
650    _2
$a nádorové buněčné linie $7 D045744
655    _2
$a časopisecké články $7 D016428
655    _2
$a práce podpořená grantem $7 D013485
700    1_
$a Jansen, Séverine $u Central European Institute of Technology, Masaryk University, Brno, Czech Republic
700    1_
$a Narasimhan, Subhash $u National Centre for Biomolecular Research, Faculty of Science, Masaryk University, Brno, Czech Republic; Central European Institute of Technology, Masaryk University, Brno, Czech Republic
700    1_
$a Laníková, Alice $u National Centre for Biomolecular Research, Faculty of Science, Masaryk University, Brno, Czech Republic; Central European Institute of Technology, Masaryk University, Brno, Czech Republic
700    1_
$a Lewitzky, Marc $u Institute of Molecular Medicine, Martin-Luther-University Halle-Wittenberg, Halle, Germany
700    1_
$a Feller, Stephan M $u Institute of Molecular Medicine, Martin-Luther-University Halle-Wittenberg, Halle, Germany
700    1_
$a Žídek, Lukáš $u National Centre for Biomolecular Research, Faculty of Science, Masaryk University, Brno, Czech Republic; Central European Institute of Technology, Masaryk University, Brno, Czech Republic. Electronic address: lzidek@chemi.muni.cz
773    0_
$w MED00002546 $t The Journal of biological chemistry $x 1083-351X $g Roč. 298, č. 10 (2022), s. 102384
856    41
$u https://pubmed.ncbi.nlm.nih.gov/35987383 $y Pubmed
910    __
$a ABA008 $b sig $c sign $y p $z 0
990    __
$a 20230120 $b ABA008
991    __
$a 20230131150637 $b ABA008
999    __
$a ok $b bmc $g 1891758 $s 1184515
BAS    __
$a 3
BAS    __
$a PreBMC-MEDLINE
BMC    __
$a 2022 $b 298 $c 10 $d 102384 $e 20220817 $i 1083-351X $m The Journal of biological chemistry $n J Biol Chem $x MED00002546
LZP    __
$a Pubmed-20230120

Najít záznam

Citační ukazatele

Pouze přihlášení uživatelé

Možnosti archivace