SASDAV5: apo XMRV RT (XM)

基本情報

• 登録情報: データベース: SASBDB / ID: SASDAV5

試料

apo XMRV RT

• apo XMRV RT (protein), XM, Escherichia coli

• 生物種: Escherichia coli (大腸菌)
• 引用: ジャーナル: Nucleic Acids Res / 年: 2013; タイトル: Structural analysis of monomeric retroviral reverse transcriptase in complex with an RNA/DNA hybrid.; 著者: Elzbieta Nowak / Wojciech Potrzebowski / Petr V Konarev / Jason W Rausch / Marion K Bona / Dmitri I Svergun / Janusz M Bujnicki / Stuart F J Le Grice / Marcin Nowotny / ; 要旨: A key step in proliferation of retroviruses is the conversion of their RNA genome to double-stranded DNA, a process catalysed by multifunctional reverse transcriptases (RTs). Dimeric and monomeric RTs have been described, the latter exemplified by the enzyme of Moloney murine leukaemia virus. However, structural information is lacking that describes the substrate binding mechanism for a monomeric RT. We report here the first crystal structure of a complex between an RNA/DNA hybrid substrate and polymerase-connection fragment of the single-subunit RT from xenotropic murine leukaemia virus-related virus, a close relative of Moloney murine leukaemia virus. A comparison with p66/p51 human immunodeficiency virus-1 RT shows that substrate binding around the polymerase active site is conserved but differs in the thumb and connection subdomains. Small-angle X-ray scattering was used to model full-length xenotropic murine leukaemia virus-related virus RT, demonstrating that its mobile RNase H domain becomes ordered in the presence of a substrate-a key difference between monomeric and dimeric RTs.

登録者

• Petr Konarev (EMBL-Hamburg, European Molecular Biology Laboratory (EMBL) - Hamburg outstation, Notkestraße 85, Geb. 25A, 22607 Hamburg, Deutschland, Germany)

モデル

• モデル #141: ; タイプ: atomic / ソフトウェア: CRYSOL / カイ2乗値: 1.258884; 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: apo XMRV RT / Sample MW: 75 kDa
• バッファ: 名称: 10mM HEPES / pH: 6.5 / 組成: KCl 100.000 mM, glycerol 5.000 %

要素 #103

名称: XM / タイプ: protein / 記述: apo XMRV RT / 分子量: 75 / 分子数: 1 / 由来: Escherichia coli
配列:

MSHHHHHHSA TLNIEDEYRL HETSKEPDVP LGSTWLSDFP QAWAETGGMG LAVRQAPLII PLKATSTPVS IKQYPMSQEA RLGIKPHIQR LLDQGILVPC QSPWNTPLLP VKKPGTNDYR PVQDLREVNK RVEDIHPTVP NPYNLLSGLP PSHQWYTVLD LKDAFFCLRL HPTSQPLFAF EWRDPEMGIS GQLTWTRLPQ GFKNSPTLFD EALHRDLADF RIQHPDLILL QYVDDLLLAA TSEQDCQRGT RALLQTLGNL GYRASAKKAQ ICQKQVKYLG YLLKEGQRWL TEARKETVMG QPTPKTPRQL REFLGTAGFC RLWIPGFAEM AAPLYPLTKT GTLFNWGPDQ QKAYQEIKQA LLTAPALGLP DLTKPFELFV DEKQGYAKGV LTQKLGPWRR PVAYLSKKLD PVAAGWPPCL RMVAAIAVLT KDAGKLTMGQ PLVILAPHAV EALVKQPPDR WLSNARMTHY QAMLLDTDRV QFGPVVALNP ATLLPLPEKE APHDCLEILA ETHGTRPDLT DQPIPDADYT WYTDGSSFLQ EGQRRAGAAV TTETEVIWAR ALPAGTSAQR AELIALTQAL KMAEGKKLNV YTDSRYAFAT AHVHGEIYRR RGLLTSEGRE IKNKNEILAL LKALFLPKRL SIIHCPGHQK GNSAEARGNR MADQAAREAA MKAVLETSTL L

実験情報

• ビーム: 設備名称: DORIS III X33 / 地域: Hamburg / 国: Germany / 形状: 0.6 / 線源: X-ray synchrotron / 波長: 0.15 Å / スペクトロメータ・検出器間距離: 2.7 mm
• 検出器: 名称: Pilatus 1M-W / Pixsize x: 0.172 mm

スキャン

タイトル: apo XMRV RT / 測定日: 2011年12月8日 / 保管温度: 10 °C / セル温度: 10 °C / 照射時間: 15 sec. / フレーム数: 8 / 単位: 1/nm /

	Min	Max
Q	0.0855	6.032

距離分布関数 P(R)

ソフトウェア P(R): GNOM 4.5a / ポイント数: 443 /

	Min	Max
Q	0.1526	2.578
P(R) point	25	467
R	0	13.57

結果

D max: 13.5 / カーブのタイプ: single_conc /

	実験値	Porod
分子量	75.5 kDa	-
体積	-	160 nm3

	P(R)	Guinier
前方散乱 I0	45	45.31
慣性半径, Rg	4.01 nm	4 nm