SASDB44: Aureochrome 1a from P. tricornutum, amino acids 148-378 (N-termin...

基本情報

• 登録情報: データベース: SASBDB / ID: SASDB44

試料

Aureochrome 1a from P. tricornutum, amino acids 148-378 (N-terminal truncation), dark state

• Aureochrome 1a (N-terminally truncated) (protein), Phaeodactylum tricornutum

• 生物種: Phaeodactylum tricornutum (海洋性珪藻)
• 引用: ジャーナル: Elife / 年: 2016; タイトル: Blue light-induced LOV domain dimerization enhances the affinity of Aureochrome 1a for its target DNA sequence.; 著者: Udo Heintz / Ilme Schlichting / ; 要旨: The design of synthetic optogenetic tools that allow precise spatiotemporal control of biological processes previously inaccessible to optogenetic control has developed rapidly over the last years. Rational design of such tools requires detailed knowledge of allosteric light signaling in natural photoreceptors. To understand allosteric communication between sensor and effector domains, characterization of all relevant signaling states is required. Here, we describe the mechanism of light-dependent DNA binding of the light-oxygen-voltage (LOV) transcription factor Aureochrome 1a from Phaeodactylum tricornutum (PtAu1a) and present crystal structures of a dark state LOV monomer and a fully light-adapted LOV dimer. In combination with hydrogen/deuterium-exchange, solution scattering data and DNA-binding experiments, our studies reveal a light-sensitive interaction between the LOV and basic region leucine zipper DNA-binding domain that together with LOV dimerization results in modulation of the DNA affinity of PtAu1a. We discuss the implications of these results for the design of synthetic LOV-based photosensors with application in optogenetics.

登録者

• Udo Heintz (Max Planck Institute for Medical Research, Germany)

モデル

• モデル #481: ; タイプ: dummy / ソフトウェア: DAMSTART/DAMMIN / ダミー原子の半径: 2.50 A / 対称性: P1 / カイ2乗値: 8.567329; Omokage検索でこの集合体の類似形状データを探す (詳細)

試料

• 試料: 名称: Aureochrome 1a from P. tricornutum, amino acids 148-378 (N-terminal truncation), dark state
• バッファ: 名称: HEPES / 濃度: 20.00 mM / pH: 7.5 / 組成: 100 mM NaCl, 10 mM MgCl2, 5% w/v glycerol

要素 #313

タイプ: protein / 記述: Aureochrome 1a (N-terminally truncated) / 分子量: 26.277 / 分子数: 2 / 由来: Phaeodactylum tricornutum
配列:

GAMGMSEQQK VERRERNREH AKRSRIRKKF LLESLQQSVS LLKEENEKLK TSIRSHLGEE KADTLIDSAN NNKTDVDGLL ASSQGIANKV LDDPDFSFIK ALQTAQQNFV VTDPSLPDNP IVYASQGFLN LTGYSLDQIL GRNCRFLQGP ETDPKAVERI RKAIEQGNDM SVCLLNYRVD GTTFWNQFFI AALRDAGGNV TNFVGVQCKV SDQYAATVTK QQEEEEEAAA NDDED

実験情報

• ビーム: 設備名称: Swiss Light Source cSAXS / 地域: Villigen / 国: Switzerland / 線源: X-ray synchrotron / 波長: 0.1 Å / スペクトロメータ・検出器間距離: 2.15 mm
• 検出器: 名称: Pilatus 2M

スキャン

タイトル: N-terminally truncated Aureochrome 1a / 測定日: 2015年3月11日 / 保管温度: 10 °C / セル温度: 10 °C / 照射時間: 0.5 sec. / フレーム数: 10 / 単位: 1/nm /

	Min	Max
Q	0.1419	3.2007

距離分布関数 P(R)

ソフトウェア P(R): GNOM 4.5a / ポイント数: 282 /

	Min	Max
Q	0.01671	0.2735
P(R) point	5	286
R	0	97.9

結果

カーブのタイプ: single_conc

	実験値	Porod	Estimated	Estimated method
分子量	54.225 kDa	56.403 kDa	-	-
体積	-	90.25 nm3	108.5	DAMMIN ab initio model

	P(R)	P(R) error	Guinier	Guinier error
前方散乱 I0	57.17	0.03	58.02	0.06
慣性半径, Rg	2.804 nm	0.002	2.88 nm	0.01

	Min	Max
D	-	9.79
Guinier point	1	60