PLUCAROVÁ, Jitka, Séverine JANSEN, Subhash NARASIMHAN, Alice LANÍKOVÁ, Marc LEWITZKY, M. Stephan FELLER and Lukáš ŽÍDEK. Specific phosphorylation of microtubule-associated protein 2c by extracellular signal–regulated kinase reduces interactions at its Pro-rich regions. JOURNAL OF BIOLOGICAL CHEMISTRY. UNITED STATES: ELSEVIER, vol. 298, No 10, p. 102384-102399. ISSN 0021-9258. doi:10.1016/j.jbc.2022.102384. 2022.
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Basic information
Original name Specific phosphorylation of microtubule-associated protein 2c by extracellular signal–regulated kinase reduces interactions at its Pro-rich regions
Authors PLUCAROVÁ, Jitka (203 Czech Republic, belonging to the institution), Séverine JANSEN (250 France, belonging to the institution), Subhash NARASIMHAN (356 India, belonging to the institution), Alice LANÍKOVÁ (203 Czech Republic, belonging to the institution), Marc LEWITZKY, M. Stephan FELLER and Lukáš ŽÍDEK (203 Czech Republic, guarantor, belonging to the institution).
Edition JOURNAL OF BIOLOGICAL CHEMISTRY, UNITED STATES, ELSEVIER, 2022, 0021-9258.
Other information
Original language English
Type of outcome Article in a journal
Field of Study 10608 Biochemistry and molecular biology
Country of publisher United States of America
Confidentiality degree is not subject to a state or trade secret
WWW URL
Impact factor Impact factor: 5.157 in 2020
RIV identification code RIV/00216224:14740/22:00126868
Organization unit Central European Institute of Technology
Doi http://dx.doi.org/10.1016/j.jbc.2022.102384
UT WoS 000867425500004
Keywords in English NMR; PKA; Src homology 3 domain; cyclin-dependent kinase; extracellular signal–regulated kinase; growth factor receptor-bound protein 2 (GRB2); microtubule-associated protein
Tags CF BIC, CF NMR, CF PROT, rivok
Tags International impact, Reviewed
Changed by Changed by: prof. Mgr. Lukáš Žídek, Ph.D., učo 38990. Changed: 30/1/2023 10:12.
Abstract
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.
Links
GA20-12669S, research and development projectName: Interakce určující fyziologické funkce proteinu Microtubule Associated Protein 2c s atomovým rozlišením
Investor: Czech Science Foundation
LM2018127, research and development projectName: Česká infrastruktura pro integrativní strukturní biologii (Acronym: CIISB)
Investor: Ministry of Education, Youth and Sports of the CR
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