Efficient protocol for backbone and side-chain assignments of
large, intrinsically disordered proteins: transient secondary
structure analysis of 49.2 kDa microtubule associated protein
2c

J 2013

Efficient protocol for backbone and side-chain assignments of large, intrinsically disordered proteins: transient secondary structure analysis of 49.2 kDa microtubule associated protein 2c

NOVÁČEK, Jiří, Lubomír JANDA, Radka DOPITOVÁ, Lukáš ŽÍDEK, Vladimír SKLENÁŘ et. al.

Základní údaje

Originální název

Efficient protocol for backbone and side-chain assignments of large, intrinsically disordered proteins: transient secondary structure analysis of 49.2 kDa microtubule associated protein 2c

Autoři

NOVÁČEK, Jiří (203 Česká republika, domácí), Lubomír JANDA (203 Česká republika, domácí), Radka DOPITOVÁ (203 Česká republika, domácí), Lukáš ŽÍDEK (203 Česká republika, garant, domácí) a Vladimír SKLENÁŘ (203 Česká republika, domácí)

Vydání

Journal of Biomolecular NMR, Dordrecht, Springer, 2013, 0925-2738

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10610 Biophysics

Stát vydavatele

Nizozemské království

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 3.305

Kód RIV

RIV/00216224:14740/13:00066377

Organizační jednotka

Středoevropský technologický institut

DOI

http://dx.doi.org/10.1007/s10858-013-9761-7

UT WoS

000323661800001

Klíčová slova anglicky

Nuclear magnetic resonance; Intrinsically disordered proteins; Microtubule-associated protein; Transient secondary structure; C-13 detection

Štítky

ok, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 24. 10. 2013 09:16, prof. Mgr. Lukáš Žídek, Ph.D.

Anotace

V originále

Microtubule-associated proteins (MAPs) are abundantly present in axons and dendrites, and have been shown to play crucial role during the neuronal morphogenesis. The period of main dendritic outgrowth and synaptogenesis coincides with high expression levels of one of MAPs, the MAP2c, in rats. The MAP2c is a 49.2 kDa intrinsically disordered protein. To achieve an atomic resolution characterization of such a large protein, we have developed a protocol based on the acquisition of two five-dimensional C-13-directly detected NMR experiments. Our previously published 5D CACONCACO experiment (Novacek et al. in J Biomol NMR 50(1):1-11, 2011) provides the sequential assignment of the backbone resonances, which is not interrupted by the presence of the proline residues in the amino acid sequence. A novel 5D HC(CC-TOCSY)CACON experiment facilitates the assignment of the aliphatic side chain resonances. To streamline the data analysis, we have developed a semi-automated procedure for signal assignments. The obtained data provides the first atomic resolution insight into the conformational state of MAP2c and constitutes a model for further functional studies of MAPs.

Návaznosti

ED1.1.00/02.0068, projekt VaV

Název: CEITEC - central european institute of technology

GAP206/11/0758, projekt VaV

Název: Vývoj metodologie s vysokým rozlišením pro NMR studie neuspořádaných proteinů s vysoce degenerovanými rezonančními frekvencemi (Akronym: HIGHRES)

Investor: Grantová agentura ČR, Vývoj metodologie s vysokým rozlišením pro NMR studie neuspořádaných proteinů s vysoce degenerovanými rezonančními frekvencemi

261863, interní kód MU

Název: BioNMR Facilities - Bio NMR (Akronym: BioNMR)

Investor: Evropská unie, BioNMR Facilities - Bio NMR, Kapacity

Citovat

NOVÁČEK, Jiří, Lubomír JANDA, Radka DOPITOVÁ, Lukáš ŽÍDEK a Vladimír SKLENÁŘ. Efficient protocol for backbone and side-chain assignments of large, intrinsically disordered proteins: transient secondary structure analysis of 49.2 kDa microtubule associated protein 2c. Journal of Biomolecular NMR. Dordrecht: Springer, 2013, roč. 56, č. 4, s. 291-301. ISSN 0925-2738. Dostupné z: https://dx.doi.org/10.1007/s10858-013-9761-7.

@article{1123258,
   author = {Nováček, Jiří and Janda, Lubomír and Dopitová, Radka and Žídek, Lukáš and Sklenář, Vladimír},
   article_location = {Dordrecht},
   article_number = {4},
   doi = {http://dx.doi.org/10.1007/s10858-013-9761-7},
   keywords = {Nuclear magnetic resonance; Intrinsically disordered proteins; Microtubule-associated protein; Transient secondary structure; C-13 detection},
   language = {eng},
   issn = {0925-2738},
   journal = {Journal of Biomolecular NMR},
   title = {Efficient protocol for backbone and side-chain assignments of large, intrinsically disordered proteins: transient secondary structure analysis of 49.2 kDa microtubule associated protein 2c},
   url = {http://www.ncbi.nlm.nih.gov/pubmed/23877929},
   volume = {56},
   year = {2013}
}

TY  - JOUR
ID  - 1123258
AU  - Nováček, Jiří - Janda, Lubomír - Dopitová, Radka - Žídek, Lukáš - Sklenář, Vladimír
PY  - 2013
TI  - Efficient protocol for backbone and side-chain assignments of large, intrinsically disordered proteins: transient secondary structure analysis of 49.2 kDa microtubule associated protein 2c
JF  - Journal of Biomolecular NMR
VL  - 56
IS  - 4
SP  - 291-301
EP  - 291-301
PB  - Springer
SN  - 09252738
KW  - Nuclear magnetic resonance
KW  - Intrinsically disordered proteins
KW  - Microtubule-associated protein
KW  - Transient secondary structure
KW  - C-13 detection
UR  - http://www.ncbi.nlm.nih.gov/pubmed/23877929
L2  - http://www.ncbi.nlm.nih.gov/pubmed/23877929
N2  - Microtubule-associated proteins (MAPs) are abundantly present in axons and dendrites, and have been shown to play crucial role during the neuronal morphogenesis. The period of main dendritic outgrowth and synaptogenesis coincides with high expression levels of one of MAPs, the MAP2c, in rats. The MAP2c is a 49.2 kDa intrinsically disordered protein. To achieve an atomic resolution characterization of such a large protein, we have developed a protocol based on the acquisition of two five-dimensional C-13-directly detected NMR experiments. Our previously published 5D CACONCACO experiment (Novacek et al. in J Biomol NMR 50(1):1-11, 2011) provides the sequential assignment of the backbone resonances, which is not interrupted by the presence of the proline residues in the amino acid sequence. A novel 5D HC(CC-TOCSY)CACON experiment facilitates the assignment of the aliphatic side chain resonances. To streamline the data analysis, we have developed a semi-automated procedure for signal assignments. The obtained data provides the first atomic resolution insight into the conformational state of MAP2c and constitutes a model for further functional studies of MAPs.
ER  -

NOVÁČEK, Jiří, Lubomír JANDA, Radka DOPITOVÁ, Lukáš ŽÍDEK a Vladimír SKLENÁŘ. Efficient protocol for backbone and side-chain assignments of large, intrinsically disordered proteins: transient secondary structure analysis of 49.2 kDa microtubule associated protein 2c. \textit{Journal of Biomolecular NMR}. Dordrecht: Springer, 2013, roč.~56, č.~4, s.~291-301. ISSN~0925-2738. Dostupné z: https://dx.doi.org/10.1007/s10858-013-9761-7.

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