SASDC53: Colicin N Translocation domain (ColN-T)

基本情報

• 登録情報: データベース: SASBDB / ID: SASDC53

試料

Colicin N Translocation domain

• Colicin N Translocation domain (protein), ColN-T, Escherichia coli

機能・相同性

分子機能:

negative regulation of ion transmembrane transporter activity / pore-forming activity / defense response to Gram-negative bacterium / transmembrane transporter binding / killing of cells of another organism / plasma membrane

ドメイン・相同性:

Channel forming colicin, C-terminal cytotoxic / Channel forming colicin, C-terminal domain superfamily / Colicin pore forming domain / Channel forming colicins signature.

構成要素:

Colicin-N

• 生物種: Escherichia coli (大腸菌)
• 引用: ジャーナル: Biophys J / 年: 2017; タイトル: The Two-State Prehensile Tail of the Antibacterial Toxin Colicin N.; 著者: Christopher L Johnson / Alexandra S Solovyova / Olli Hecht / Colin Macdonald / Helen Waller / J Günter Grossmann / Geoffrey R Moore / Jeremy H Lakey / ; 要旨: Intrinsically disordered regions within proteins are critical elements in many biomolecular interactions and signaling pathways. Antibacterial toxins of the colicin family, which could provide new antibiotic functions against resistant bacteria, contain disordered N-terminal translocation domains (T-domains) that are essential for receptor binding and the penetration of the Escherichia coli outer membrane. Here we investigate the conformational behavior of the T-domain of colicin N (ColN-T) to understand why such domains are widespread in toxins that target Gram-negative bacteria. Like some other intrinsically disordered proteins in the solution state of the protein, ColN-T shows dual recognition, initially interacting with other domains of the same colicin N molecule and later, during cell killing, binding to two different receptors, OmpF and TolA, in the target bacterium. ColN-T is invisible in the high-resolution x-ray model and yet accounts for 90 of the toxin's 387 amino acid residues. To reveal its solution structure that underlies such a dynamic and complex system, we carried out mutagenic, biochemical, hydrodynamic and structural studies using analytical ultracentrifugation, NMR, and small-angle x-ray scattering on full-length ColN and its fragments. The structure was accurately modeled from small-angle x-ray scattering data by treating ColN as a flexible system, namely by the ensemble optimization method, which enables a distribution of conformations to be included in the final model. The results reveal, to our knowledge, for the first time the dynamic structure of a colicin T-domain. ColN-T is in dynamic equilibrium between a compact form, showing specific self-recognition and resistance to proteolysis, and an extended form, which most likely allows for effective receptor binding.

登録者

• Alex Solovyova (Newcastle University, Newcastle, UK)

モデル

• モデル #1211: ; タイプ: atomic / ダミー原子の半径: 1.90 A / カイ2乗値: 1.507984 / P-value: 0.000494; 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 #1212: ; タイプ: atomic / ダミー原子の半径: 1.90 A / カイ2乗値: 1.507984 / P-value: 0.000494; 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: Colicin N Translocation domain / 試料濃度: 0.40-6.30
• バッファ: 名称: 50 mM Na-Phosphate 300 mM NaCl / pH: 7.6

要素 #627

名称: ColN-T / タイプ: protein / 記述: Colicin N Translocation domain / 分子量: 9.965 / 分子数: 1 / 由来: Escherichia coli / 参照: UniProt: P08083
配列:

MGSNGADNAH NNAFGGGKNP GIGNTSGAGS NGSASSNRGN SNGWSWSNKP HKNDGFHSDG SYHITFHGDN NSKPKPGGNS GNRGNNGDGA SSHHHHHH

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: 0.15 Å / スペクトロメータ・検出器間距離: 2.85 mm
• 検出器: 名称: Pilatus 1M / タイプ: Dectris / Pixsize x: 172 mm

スキャン

タイトル: Colicin N Translocation domain / 測定日: 2012年6月29日 / 保管温度: 4 °C / セル温度: 4 °C / 照射時間: 2 sec. / フレーム数: 10 / 単位: 1/nm /

	Min	Max
Q	0.032	4.5484

距離分布関数 P(R)

ソフトウェア P(R): GNOM 5.0 / ポイント数: 948 /

	Min	Max
Q	0.22759	3.78483
P(R) point	1	948
R	0	11.4

結果

カーブのタイプ: single_conc
コメント: The results obtained from the bayesapp estimation of distribution functions from small-angle scattering data (http://www.bayesapp.org/) are included in the full entry zip archive.

	実験値	Standard	Standard error	Porod
分子量	10 kDa	18.5 kDa	0.88	-
体積	-	-	-	21.82 nm3

	P(R)	P(R) error	Guinier	Guinier error
前方散乱 I0	23.4	0.0777	22.95	0.044
慣性半径, Rg	2.93 nm	0.02	2.76 nm	0.15

	Min	Max	Error
D	-	11.42	0.25
Guinier point	53	110	-