SASDBL7: N-terminal domain of Vaccinia virus A46 protein (1-83)

基本情報

• 登録情報: データベース: SASBDB / ID: SASDBL7

試料

N-terminal domain of Vaccinia virus A46 protein (1-83)

• Protein A46 (protein), A46-delta, Vaccinia virus

• 機能・相同性: Poxvirus Bcl-2-like / Poxvirus domain superfamily / Poxvirus Bcl-2-like proteins / extrinsic component of cytoplasmic side of plasma membrane / protein sequestering activity / symbiont-mediated suppression of host toll-like receptor signaling pathway / symbiont-mediated suppression of host cytoplasmic pattern recognition receptor signaling pathway via inhibition of IRF3 activity / symbiont-mediated suppression of host NF-kappaB cascade / Protein OPG176
• 生物種: Vaccinia virus (ウイルス)
• 引用: ジャーナル: PLoS Pathog / 年: 2016; タイトル: Vaccinia Virus Immunomodulator A46: A Lipid and Protein-Binding Scaffold for Sequestering Host TIR-Domain Proteins.; 著者: Sofiya Fedosyuk / Gustavo Arruda Bezerra / Katharina Radakovics / Terry K Smith / Massimo Sammito / Nina Bobik / Adam Round / Lynn F Ten Eyck / Kristina Djinović-Carugo / Isabel Usón / Tim Skern / ; 要旨: Vaccinia virus interferes with early events of the activation pathway of the transcriptional factor NF-kB by binding to numerous host TIR-domain containing adaptor proteins. We have previously determined the X-ray structure of the A46 C-terminal domain; however, the structure and function of the A46 N-terminal domain and its relationship to the C-terminal domain have remained unclear. Here, we biophysically characterize residues 1-83 of the N-terminal domain of A46 and present the X-ray structure at 1.55 Å. Crystallographic phases were obtained by a recently developed ab initio method entitled ARCIMBOLDO_BORGES that employs tertiary structure libraries extracted from the Protein Data Bank; data analysis revealed an all β-sheet structure. This is the first such structure solved by this method which should be applicable to any protein composed entirely of β-sheets. The A46(1-83) structure itself is a β-sandwich containing a co-purified molecule of myristic acid inside a hydrophobic pocket and represents a previously unknown lipid-binding fold. Mass spectrometry analysis confirmed the presence of long-chain fatty acids in both N-terminal and full-length A46; mutation of the hydrophobic pocket reduced the lipid content. Using a combination of high resolution X-ray structures of the N- and C-terminal domains and SAXS analysis of full-length protein A46(1-240), we present here a structural model of A46 in a tetrameric assembly. Integrating affinity measurements and structural data, we propose how A46 simultaneously interferes with several TIR-domain containing proteins to inhibit NF-κB activation and postulate that A46 employs a bipartite binding arrangement to sequester the host immune adaptors TRAM and MyD88.

登録者

• Sofiya Fedosyuk (Medical University of Vienna)

モデル

• モデル #855: ; タイプ: mix / ソフトウェア: CORAL / ダミー原子の半径: 1.90 A / カイ2乗値: 0.657; 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: N-terminal domain of Vaccinia virus A46 protein (1-83); 試料濃度: 15.5 mg/ml
• バッファ: 名称: 20 mM Tris-HCl, 10 mM DTT / pH: 8.5

要素 #462

名称: A46-delta / タイプ: protein / 記述: Protein A46 / 分子量: 10.028 / 分子数: 4 / 由来: Vaccinia virus / 参照: UniProt: P26672
配列:

GSMAQQMAFD ISVNASKTIN ALVYFSTQQN KLVIRNEVND THYTVEFDRD KVVDTFISYN RHNDTIEIRG VLPEETNIGC AVNTPVSMT

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: 0.09918 Å / スペクトロメータ・検出器間距離: 2.867 mm
• 検出器: 名称: Pilatus 1M

スキャン

タイトル: N-terminal domain of Vaccinia virus A46 protein (1
測定日: 2015年6月25日 / セル温度: 20 °C / 照射時間: 0.1 sec. / 単位: 1/nm /

	Min	Max
Q	0.1315	4.9336

距離分布関数 P(R)

ソフトウェア P(R): GNOM 5.0 / ポイント数: 378 /

	Min	Max
Q	0.13616	1.90933
P(R) point	1	378
R	0	9

結果

カーブのタイプ: single_conc /

	実験値	Porod
分子量	38.857 kDa	38.857 kDa
体積	-	68 nm3

	P(R)	P(R) error	Guinier	Guinier error
前方散乱 I0	14.97	0.03	14.91	0.02
慣性半径, Rg	2.65 nm	0.01	2.61 nm	0.09

	Min	Max
D	-	9
Guinier point	1	65