Functionally specific binding regions of microtubule-associated
protein 2c exhibit distinct conformations and dynamics

J 2018

Functionally specific binding regions of microtubule-associated protein 2c exhibit distinct conformations and dynamics

MELKOVÁ, Kateřina; Vojtěch ZAPLETAL; Séverine JANSEN; Erik NOMILNER; Milan ZACHRDLA et al.

Základní údaje

Originální název

Functionally specific binding regions of microtubule-associated protein 2c exhibit distinct conformations and dynamics

Autoři

MELKOVÁ, Kateřina; Vojtěch ZAPLETAL; Séverine JANSEN; Erik NOMILNER; Milan ZACHRDLA; Jozef HRITZ; Jiří NOVÁČEK; M. ZWECKSTETTER; M.R. JENSEN; M. BLACKLEDGE a Lukáš ŽÍDEK

Vydání

Journal of Biological Chemistry, Bethesda, USA, Amer. Soc. Biochem. Mol. Biol. 2018, 0021-9258

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10608 Biochemistry and molecular biology

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 4.106

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:14740/18:00101098

Organizační jednotka

Středoevropský technologický institut

Klíčová slova anglicky

NMR relaxation; Tau protein (Tau); microtubule-associated protein (MAP); nuclear magnetic resonance (NMR); paramagnetic relaxation enhancement (PRE); protein conformation; small-angle X-ray scattering (SAXS)

Štítky

CF NMR, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 13. 3. 2019 13:21, Mgr. Pavla Foltynová, Ph.D.

Anotace

V originále

Microtubule-associated protein 2c (MAP2c) is a 49-kDa intrinsically disordered protein regulating the dynamics of microtubules in developing neurons. MAP2c differs from its sequence homologue Tau in the pattern and kinetics of phosphorylation by cAMP-dependent protein kinase (PKA). Moreover, the mechanisms through which MAP2c interacts with its binding partners and the conformational changes and dynamics associated with these interactions remain unclear. Here, we used NMR relaxation and paramagnetic relaxation enhancement techniques to determine the dynamics and long-range interactions within MAP2c. The relaxation rates revealed large differences in flexibility of individual regions of MAP2c, with the lowest flexibility observed in the known and proposed binding sites. Quantitative conformational analyses of chemical shifts, small-angle X-ray scattering (SAXS), and paramagnetic relaxation enhancement measurements disclosed that MAP2c regions interacting with important protein partners, including Fyn tyrosine kinase, plectin, and PKA, adopt specific conformations. High populations of polyproline II and alpha-helices were found in Fyn- and plectin-binding sites of MAP2c, respectively. The region binding the regulatory subunit of PKA consists of two helical motifs bridged by a more extended conformation. Of note, although MAP2c and Tau did not differ substantially in their conformations in regions of high sequence identity, we found that they differ significantly in long-range interactions, dynamics, and local conformation motifs in their N-terminal domains. These results highlight that the N-terminal regions of MAP2c provide important specificity to its regulatory roles and indicate a close relationship between MAP2c's biological functions and conformational behavior.

Návaznosti

EF16_013/0001776, projekt VaV

Název: Česká infrastruktura pro integrativní strukturní biologii pro lidské zdraví

GA15-14974S, projekt VaV

Název: Charakterizace proteinu MAP2c (microtubule associated protein 2c) a modifikací regulujících jeho funkci s atomovým rozlišením

Investor: Grantová agentura ČR, Charakterizace proteinu MAP2c (microtubule associated protein 2c) a modifikací regulujících jeho funkci s atomovým rozlišením

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

LM2015085, projekt VaV

Název: CERIT Scientific Cloud (Akronym: CERIT-SC)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, CERIT Scientific Cloud

692068, interní kód MU

Název: BISON - Bridging Structural Biology with Biological Synthesis and Self Assembly to Reveal Key Processes in Living Systems (Akronym: BISON)

Investor: Evropská unie, BISON - Bridging Structural Biology with Biological Synthesis and Self Assembly to Reveal Key Processes in Living Systems, Spreading excellence and widening participation

Přiložené soubory

melkova-revised.pdf Verze souboru

Citovat

MELKOVÁ, Kateřina; Vojtěch ZAPLETAL; Séverine JANSEN; Erik NOMILNER; Milan ZACHRDLA; Jozef HRITZ; Jiří NOVÁČEK; M. ZWECKSTETTER; M.R. JENSEN; M. BLACKLEDGE a Lukáš ŽÍDEK. Functionally specific binding regions of microtubule-associated protein 2c exhibit distinct conformations and dynamics. Journal of Biological Chemistry. Bethesda, USA: Amer. Soc. Biochem. Mol. Biol., 2018, roč. 293, č. 34, s. 13297-13309. ISSN 0021-9258. Dostupné z: https://doi.org/10.1074/jbc.RA118.001769.

@article{1427578,
   author = {Melková, Kateřina and Zapletal, Vojtěch and Jansen, Séverine and Nomilner, Erik and Zachrdla, Milan and Hritz, Jozef and Nováček, Jiří and Zweckstetter, M. and Jensen, M.R. and Blackledge, M. and Žídek, Lukáš},
   article_location = {Bethesda, USA},
   article_number = {34},
   doi = {https://doi.org/10.1074/jbc.RA118.001769},
   keywords = {NMR relaxation; Tau protein (Tau); microtubule-associated protein (MAP); nuclear magnetic resonance (NMR); paramagnetic relaxation enhancement (PRE); protein conformation; small-angle X-ray scattering (SAXS)},
   language = {eng},
   issn = {0021-9258},
   journal = {Journal of Biological Chemistry},
   title = {Functionally specific binding regions of microtubule-associated protein 2c exhibit distinct conformations and dynamics},
   url = {http://www.jbc.org/content/early/2018/06/20/jbc.RA118.001769.abstract},
   volume = {293},
   year = {2018}
}

TY  - JOUR
ID  - 1427578
AU  - Melková, Kateřina - Zapletal, Vojtěch - Jansen, Séverine - Nomilner, Erik - Zachrdla, Milan - Hritz, Jozef - Nováček, Jiří - Zweckstetter, M. - Jensen, M.R. - Blackledge, M. - Žídek, Lukáš
PY  - 2018
TI  - Functionally specific binding regions of microtubule-associated protein 2c exhibit distinct conformations and dynamics
JF  - Journal of Biological Chemistry
VL  - 293
IS  - 34
SP  - 13297-13309
EP  - 13297-13309
PB  - Amer. Soc. Biochem. Mol. Biol.
SN  - 00219258
KW  - NMR relaxation
KW  - Tau protein (Tau)
KW  - microtubule-associated protein (MAP)
KW  - nuclear magnetic resonance (NMR)
KW  - paramagnetic relaxation enhancement (PRE)
KW  - protein conformation
KW  - small-angle X-ray scattering (SAXS)
UR  - http://www.jbc.org/content/early/2018/06/20/jbc.RA118.001769.abstract
N2  - Microtubule-associated protein 2c (MAP2c) is a 49-kDa intrinsically disordered protein regulating the dynamics of microtubules in developing neurons. MAP2c differs from its sequence homologue Tau in the pattern and kinetics of phosphorylation by cAMP-dependent protein kinase (PKA). Moreover, the mechanisms through which MAP2c interacts with its binding partners and the conformational changes and dynamics associated with these interactions remain unclear. Here, we used NMR relaxation and paramagnetic relaxation enhancement techniques to determine the dynamics and long-range interactions within MAP2c. The relaxation rates revealed large differences in flexibility of individual regions of MAP2c, with the lowest flexibility observed in the known and proposed binding sites. Quantitative conformational analyses of chemical shifts, small-angle X-ray scattering (SAXS), and paramagnetic relaxation enhancement measurements disclosed that MAP2c regions interacting with important protein partners, including Fyn tyrosine kinase, plectin, and PKA, adopt specific conformations. High populations of polyproline II and alpha-helices were found in Fyn- and plectin-binding sites of MAP2c, respectively. The region binding the regulatory subunit of PKA consists of two helical motifs bridged by a more extended conformation. Of note, although MAP2c and Tau did not differ substantially in their conformations in regions of high sequence identity, we found that they differ significantly in long-range interactions, dynamics, and local conformation motifs in their N-terminal domains. These results highlight that the N-terminal regions of MAP2c provide important specificity to its regulatory roles and indicate a close relationship between MAP2c's biological functions and conformational behavior.
ER  -

MELKOVÁ, Kateřina; Vojtěch ZAPLETAL; Séverine JANSEN; Erik NOMILNER; Milan ZACHRDLA; Jozef HRITZ; Jiří NOVÁČEK; M. ZWECKSTETTER; M.R. JENSEN; M. BLACKLEDGE a Lukáš ŽÍDEK. Functionally specific binding regions of microtubule-associated protein 2c exhibit distinct conformations and dynamics. \textit{Journal of Biological Chemistry}. Bethesda, USA: Amer. Soc. Biochem. Mol. Biol., 2018, roč.~293, č.~34, s.~13297-13309. ISSN~0021-9258. Dostupné z: https://doi.org/10.1074/jbc.RA118.001769.

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