SASDD62: Microtubule associated protein MAP2c (isoform 3); 12 mg/ml

基本情報

登録情報

データベース: SASBDB / ID: SASDD62

試料

Microtubule associated protein MAP2c (isoform 3); 12 mg/ml

• Microtubule-associated protein 2, isoform 3 (protein), MAP2c, Rattus norvegicus

• 機能・相同性: Isoform 3 of Microtubule-associated protein 2
• 生物種: Rattus norvegicus (ドブネズミ)
• 引用: ジャーナル: J Biol Chem / 年: 2018; タイトル: Functionally specific binding regions of microtubule-associated protein 2c exhibit distinct conformations and dynamics.; 著者: Kateřina Melková / Vojtěch Zapletal / Séverine Jansen / Erik Nomilner / Milan Zachrdla / Jozef Hritz / Jiří Nováček / Markus Zweckstetter / Malene R Jensen / Martin Blackledge / Lukáš Žídek / ; 要旨: 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 α-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.

登録者

• Vojtech Zapletal (MUNI, Masaryk University; Research group Structure and Dynamics of Proteins., Brno, Czech Republic)

モデル

試料

• 試料: 名称: Microtubule associated protein MAP2c (isoform 3); 12 mg/ml; 試料濃度: 12 mg/ml
• バッファ: 名称: 50 mM MOPS, 150 mM NaCl, 0.03% NaN3 / pH: 6.9 / コメント: aka: MOPS map2c buffer

要素 #917

名称: MAP2c / タイプ: protein / 記述: Microtubule-associated protein 2, isoform 3 / 分子量: 49.299 / 分子数: 1 / 由来: Rattus norvegicus / 参照: UniProt: P15146-3
配列:

MADERKDEGK APHWTSASLT EAAAHPHSPE MKDQGGSGEG LSRSANGFPY REEEEGAFGE HGSQGTYSDT KENGINGELT SADRETAEEV SARIVQVVTA EAVAVLKGEQ EKEAQHKDQP AALPLAAEET VNLPPSPPPS PASEQTAALE EATSGESAQA PSAFKQAKDK VTDGITKSPE KRSSLPRPSS ILPPRRGVSG DREENSFSLN SSISSARRTT RSEPIRRAGK SGTSTPTTPG STAITPGTPP SYSSRTPGTP GTPSYPRTPG TPKSGILVPS EKKVAIIRTP PKSPATPKQL RLINQPLPDL KNVKSKIGST DNIKYQPKGG QVQIVTKKID LSHVTSKCGS LKNIRHRPGG GRVKIESVKL DFKEKAQAKV GSLDNAHHVP GGGNVKIDSQ KLNFREHAKA RVDHGAEIIT QSPSRSSVAS PRRLSNVSSS GSINLLESPQ LATLAEDVTA ALAKQGL

実験情報

• ビーム: 設備名称: ESRF BM29 / 地域: Grenoble / 国: France / 線源: X-ray synchrotron / 波長: 0.099 Å
• 検出器: 名称: Pilatus 1M / タイプ: Dectris / Pixsize x: 172 mm

スキャン

タイトル: Microtubule associated protein MAP2c (isoform 3); 12 mg/ml
測定日: 2017年4月21日 / 保管温度: 20 °C / セル温度: 20 °C / 照射時間: 1 sec. / フレーム数: 10 / 単位: 1/nm /

	Min	Max
Q	0.0308	4.9444

結果

Experimental MW: 66 kDa / カーブのタイプ: single_conc

	Guinier	Guinier error
前方散乱 I0	63.72	0.25
慣性半径, Rg	8.29 nm	0.4

	Min	Max
Guinier point	14	24