SASDAB9: EcPaaA2-HisEcParE2 construct (Plasmid stabilization protein ParE,...

基本情報

• 登録情報: データベース: SASBDB / ID: SASDAB9

試料

EcPaaA2-HisEcParE2 construct

• Plasmid stabilization protein ParE (protein), EcParE2, Escherichia coli
• Uncharacterized protein (Antitoxin) (protein), EcPaaA2, Escherichia coli

• 機能・相同性: ParE toxin of type II toxin-antitoxin system, parDE / Toxin-antitoxin system, RelE/ParE toxin family / Toxin-antitoxin system, RelE/ParE toxin domain superfamily / Plasmid stabilization protein ParE / :
• 生物種: Escherichia coli (大腸菌)
• 引用: ジャーナル: J Mol Biol / 年: 2016; タイトル: A unique hetero-hexadecameric architecture displayed by the Escherichia coli O157 PaaA2-ParE2 antitoxin-toxin complex.; 著者: Yann G-J Sterckx / Thomas Jové / Alexander V Shkumatov / Abel Garcia-Pino / Lieselotte Geerts / Maia De Kerpel / Jurij Lah / Henri De Greve / Laurence Van Melderen / Remy Loris / ; 要旨: Many bacterial pathogens modulate their metabolic activity, virulence and pathogenicity through so-called "toxin-antitoxin" (TA) modules. The genome of the human pathogen Escherichia coli O157 contains two three-component TA modules related to the known parDE module. Here, we show that the toxin EcParE2 maps in a branch of the RelE/ParE toxin superfamily that is distinct from the branches that contain verified gyrase and ribosome inhibitors. The structure of EcParE2 closely resembles that of Caulobacter crescentus ParE but shows a distinct pattern of conserved surface residues, in agreement with its apparent inability to interact with GyrA. The antitoxin EcPaaA2 is characterized by two α-helices (H1 and H2) that serve as molecular recognition elements to wrap itself around EcParE2. Both EcPaaA2 H1 and H2 are required to sustain a high-affinity interaction with EcParE2 and for the inhibition of EcParE2-mediated killing in vivo. Furthermore, evidence demonstrates that EcPaaA2 H2, but not H1, determines specificity for EcParE2. The initially formed EcPaaA2-EcParE2 heterodimer then assembles into a hetero-hexadecamer, which is stable in solution and is formed in a highly cooperative manner. Together these findings provide novel data on quaternary structure, TA interactions and activity of a hitherto poorly characterized family of TA modules.

登録者

• Alexander Shkumatov (VUB, Vrije Universiteit Brussel, Pleinlaan 2 1050 Brussel)

モデル

• モデル #355: ; タイプ: mix / ダミー原子の半径: 1.90 A / コメント: NMA refinement / カイ2乗値: 3.439; Omokage検索でこの集合体の類似形状データを探す (詳細)
• モデル #356: ; タイプ: dummy / ダミー原子の半径: 3.50 A / カイ2乗値: 0.637; 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: EcPaaA2-HisEcParE2 construct / 試料濃度: 1.00-10.00 / Entity id: 215 / 216
• バッファ: 名称: 50 mM Tris-HCl 500 mM NaCl / 濃度: 50.00 mM / pH: 7.5 / 組成: 500 mM NaCl

要素 #215

名称: EcParE2 / タイプ: protein / 記述: Plasmid stabilization protein ParE / 分子量: 12.693 / 分子数: 8 / 由来: Escherichia coli / 参照: UniProt: A0A0D7C2L1
配列:

MGSSHHHHHH SSGLVPRGSH LPVLWLESAD TDLDDITSYI ARFDIDAAER LWQRLRGCVL PLSEHPYLYP PSDRVPGLRE IVAHPNYIIL YRVTTSSVEV VNVIHARRQF P

要素 #216

名称: EcPaaA2 / タイプ: protein / 記述: Uncharacterized protein (Antitoxin) / 分子量: 8.458 / 分子数: 8 / 由来: Escherichia coli / 参照: UniProt: A0A0F6F6Q9
配列:

MDYKDDDDKN RALSPMVSEF ETIEQENSYN EWLRAKVATS LADPRPAIPH DEVERRMAER FAKMRKERSK Q

実験情報

• ビーム: 設備名称: SOLEIL SWING / 地域: Saint-Aubin / 国: France / 線源: X-ray synchrotron / 波長: 0.1 Å / スペクトロメータ・検出器間距離: 1.82 mm
• 検出器: 名称: AVIEX / タイプ: CCD

スキャン

タイトル: parDE-like toxin-antitoxin module / 測定日: 2012年2月5日 / セル温度: 10 °C / 照射時間: 0.75 sec. / フレーム数: 750 / 単位: 1/nm /

	Min	Max
Q	0.2222	4.0026

距離分布関数 P(R)

ソフトウェア P(R): GNOM 5.0 / ポイント数: 386 /

	Min	Max
Q	0.0241114	0.206041
P(R) point	1	386
R	0	153

結果

D max: 15.28 / カーブのタイプ: merged
コメント: Use of SAXS to validate the oligomer state of the obtained crystal structure and to model missing afinity tags

	実験値	Standard	Porod
分子量	100 kDa	100 kDa	97.74 kDa
体積	-	-	166.16 nm3

	P(R)	Guinier
前方散乱 I0	0.18	0.175042
慣性半径, Rg	3.408 nm	3.255 nm