SASDA58: UL26N of pseudorabies virus (VP24, UL26N)

基本情報

• 登録情報: データベース: SASBDB / ID: SASDA58

試料

UL26N of pseudorabies virus

• VP24 (protein), UL26N, Suid herpesvirus 1

機能・相同性

分子機能:

assemblin / nuclear capsid assembly / viral release from host cell / host cell cytoplasm / serine-type endopeptidase activity / host cell nucleus / proteolysis / identical protein binding

ドメイン・相同性:

Peptidase S21 / Herpesvirus protease superfamily / Assemblin (Peptidase family S21)

構成要素:

Capsid scaffolding protein

• 生物種: Suid herpesvirus 1 (ヘルペスウイルス)
• 引用: ジャーナル: PLoS Pathog / 年: 2015; タイトル: Dimerization-Induced Allosteric Changes of the Oxyanion-Hole Loop Activate the Pseudorabies Virus Assemblin pUL26N, a Herpesvirus Serine Protease.; 著者: Martin Zühlsdorf / Sebastiaan Werten / Barbara G Klupp / Gottfried J Palm / Thomas C Mettenleiter / Winfried Hinrichs / ; 要旨: Herpesviruses encode a characteristic serine protease with a unique fold and an active site that comprises the unusual triad Ser-His-His. The protease is essential for viral replication and as such constitutes a promising drug target. In solution, a dynamic equilibrium exists between an inactive monomeric and an active dimeric form of the enzyme, which is believed to play a key regulatory role in the orchestration of proteolysis and capsid assembly. Currently available crystal structures of herpesvirus proteases correspond either to the dimeric state or to complexes with peptide mimetics that alter the dimerization interface. In contrast, the structure of the native monomeric state has remained elusive. Here, we present the three-dimensional structures of native monomeric, active dimeric, and diisopropyl fluorophosphate-inhibited dimeric protease derived from pseudorabies virus, an alphaherpesvirus of swine. These structures, solved by X-ray crystallography to respective resolutions of 2.05, 2.10 and 2.03 Å, allow a direct comparison of the main conformational states of the protease. In the dimeric form, a functional oxyanion hole is formed by a loop of 10 amino-acid residues encompassing two consecutive arginine residues (Arg136 and Arg137); both are strictly conserved throughout the herpesviruses. In the monomeric form, the top of the loop is shifted by approximately 11 Å, resulting in a complete disruption of the oxyanion hole and loss of activity. The dimerization-induced allosteric changes described here form the physical basis for the concentration-dependent activation of the protease, which is essential for proper virus replication. Small-angle X-ray scattering experiments confirmed a concentration-dependent equilibrium of monomeric and dimeric protease in solution.

登録者

• Sebastiaan Werten (Ernst-Moritz-Arndt-Universität Greifswald, Domstraße 11 17489 Greifswald)

モデル

• モデル #311: ; タイプ: atomic / ソフトウェア: Crysol / カイ2乗値: 1.288; 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 #350: ; タイプ: dummy / ソフトウェア: DAMMIN / ダミー原子の半径: 2.25 A / カイ2乗値: 0.902; 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: UL26N of pseudorabies virus / 試料濃度: 9.83 mg/ml
• バッファ: 名称: Tris/HCl / 濃度: 50.00 mM / pH: 7.5 ; 組成: 0.5 M NaCl, 0.25 M imidazole, 5% glycerol, 50 mM urea, 0.2 M MgCl2

要素 #177

名称: UL26N / タイプ: protein / 記述: VP24 / 分子量: 26.608 / 分子数: 2 / 由来: Suid herpesvirus 1 / 参照: UniProt: Q83417
配列:

MGSSHHHHHH SSGLVPRGSH MGPVYVSGYL ALYDRDGGEL ALTREIVAAA LPPAGPLPIN IDHRPRCDIG AVLAVVDDDR GPFFLGVVNC PQLGAVLARA VGPDFFGDMR LSDEERLLYL LSNYLPSASL SSRRLAPGEA PDETLFAHVA LCVIGRRVGT IVVYDASPEA AVAPFRQLSA RARSELLARA AESPDRERVW HMSEEALTRA LLSTAVNNML LRDRWELVAA RRREAGVRGH TYLQ

実験情報

• ビーム: 設備名称: PETRA III P12 / 地域: Hamburg / 国: Germany / 線源: X-ray synchrotron / 波長: 0.12 Å / スペクトロメータ・検出器間距離: 3.1 mm
• 検出器: 名称: Pilatus 2M

スキャン

タイトル: UL26N of pseudorabies virus / 測定日: 2013年9月23日 / セル温度: 10 °C / 照射時間: 0.05 sec. / フレーム数: 20 / 単位: 1/nm /

	Min	Max
Q	0.068	4.4372

距離分布関数 P(R)

ソフトウェア P(R): GNOM 5.0 / ポイント数: 905 /

	Min	Max
Q	0.096949	2.47635
P(R) point	1	905
R	0	10.56

結果

D max: 10.6 / カーブのタイプ: single_conc /

	実験値	Porod
分子量	45 kDa	45 kDa
体積	-	73 nm3

	P(R)	Guinier
前方散乱 I0	2982	2964
慣性半径, Rg	2.69 nm	2.62 nm