title: Quantitative mapping of microtubule-associated protein 2c (MAP2c) phosphorylation and regulatory protein 14-3-3ζ-binding sites reveals key differences between MAP2c and its homolog Tau.
abstract: Microtubule-associated protein 2c (MAP2c) is involved in neuronal development and is less characterized than its homolog Tau, which has various roles in neurodegeneration. Using NMR methods providing single-residue resolution and quantitative comparison, we investigated molecular interactions important for the regulatory roles of MAP2c in microtubule dynamics. We found that MAP2c and Tau significantly differ in the position and kinetics of sites that are phosphorylated by cAMP-dependent protein kinase (PKA), even in highly homologous regions. We determined the binding sites of unphosphorylated and phosphorylated MAP2c responsible for interactions with the regulatory protein 14-3-3ζ. Differences in phosphorylation and in charge distribution between MAP2c and Tau suggested that both MAP2c and Tau respond to the same signal (phosphorylation by PKA) but have different downstream effects, indicating a signaling branch point for controlling microtubule stability. Although the interactions of phosphorylated Tau with 14-3-3ζ are supposed to be a major factor in microtubule destabilization, the binding of 14-3-3ζ to MAP2c enhanced by PKA-mediated phosphorylation is likely to influence microtubule-MAP2c binding much less, in agreement with the results of our tubulin co-sedimentation measurements. The specific location of the major MAP2c phosphorylation site in a region homologous to the muscarinic receptor-binding site of Tau suggests that MAP2c also may regulate processes other than microtubule dynamics.
authors.affiliation: From the National Centre for Biomolecular Research, Faculty of Science, and.|the Central European Institute of Technology, Masaryk University, Kamenice 5, CZ-62500 Brno, Czech Republic.
authors.forename: Séverine
authors.initials: S
authors.lastname: Jansen
authors.affiliation: From the National Centre for Biomolecular Research, Faculty of Science, and.|the Central European Institute of Technology, Masaryk University, Kamenice 5, CZ-62500 Brno, Czech Republic.
authors.forename: Kateřina
authors.initials: K
authors.lastname: Melková
authors.affiliation: From the National Centre for Biomolecular Research, Faculty of Science, and.|the Central European Institute of Technology, Masaryk University, Kamenice 5, CZ-62500 Brno, Czech Republic.
authors.forename: Zuzana
authors.initials: Z
authors.lastname: Trošanová
authors.affiliation: the Central European Institute of Technology, Masaryk University, Kamenice 5, CZ-62500 Brno, Czech Republic.
authors.forename: Kateřina
authors.initials: K
authors.lastname: Hanáková
authors.affiliation: From the National Centre for Biomolecular Research, Faculty of Science, and.|the Central European Institute of Technology, Masaryk University, Kamenice 5, CZ-62500 Brno, Czech Republic.
authors.forename: Milan
authors.initials: M
authors.lastname: Zachrdla
authors.affiliation: the Central European Institute of Technology, Masaryk University, Kamenice 5, CZ-62500 Brno, Czech Republic.
authors.forename: Jiří
authors.initials: J
authors.lastname: Nováček
authors.affiliation: From the National Centre for Biomolecular Research, Faculty of Science, and.|the Central European Institute of Technology, Masaryk University, Kamenice 5, CZ-62500 Brno, Czech Republic.
authors.forename: Erik
authors.initials: E
authors.lastname: Župa
authors.affiliation: the Central European Institute of Technology, Masaryk University, Kamenice 5, CZ-62500 Brno, Czech Republic.
authors.forename: Zbyněk
authors.initials: Z
authors.lastname: Zdráhal
authors.affiliation: From the National Centre for Biomolecular Research, Faculty of Science, and jozef.hritz@ceitec.muni.cz.|the Central European Institute of Technology, Masaryk University, Kamenice 5, CZ-62500 Brno, Czech Republic.
authors.forename: Jozef
authors.initials: J
authors.lastname: Hritz
authors.affiliation: From the National Centre for Biomolecular Research, Faculty of Science, and lzidek@chemi.muni.cz.|the Central European Institute of Technology, Masaryk University, Kamenice 5, CZ-62500 Brno, Czech Republic.
authors.forename: Lukáš
authors.initials: L
authors.lastname: Žídek
mesh_terms: D048948:14-3-3 Proteins / Q000737:chemistry*; D020816:Amino Acid Motifs; D000818:Animals; D001665:Binding Sites; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D008869:Microtubule-Associated Proteins / Q000737:chemistry*; D008870:Microtubules / Q000378:metabolism; D009474:Neurons / Q000378:metabolism; D010766:Phosphorylation; D011485:Protein Binding; D051381:Rats; D015398:Signal Transduction; D014404:Tubulin / Q000378:metabolism; D016875:tau Proteins / Q000737:chemistry*
chemical_list: D048948:14-3-3 Proteins; C489787:MAP2 protein, rat; C489131:Mapt protein, rat; D008869:Microtubule-Associated Proteins; D014404:Tubulin; D016875:tau Proteins; D017868:Cyclic AMP-Dependent Protein Kinases
keywords: 14-3-3 protein; mass spectrometry (MS); microtubule-associated protein (MAP); nuclear magnetic resonance (NMR); protein kinase A (PKA)