SASDAC5: ImportinA_ImportinB (ImpA_ImpB)

基本情報

• 登録情報: データベース: SASBDB / ID: SASDAC5

試料

ImportinA_ImportinB

• ImportinA_ImportinB (protein), ImpA_ImpB, Escherichia coli

• 生物種: Escherichia coli (大腸菌)
• 引用: ジャーナル: Biochemistry / 年: 2010; タイトル: Recognition of nucleoplasmin by its nuclear transport receptor importin α/β: insights into a complete import complex.; 著者: Jorge Falces / Igor Arregi / Petr V Konarev / María A Urbaneja / Dmitri I Svergun / Stefka G Taneva / Sonia Bañuelos / ; 要旨: Nuclear import of the pentameric histone chaperone nucleoplasmin (NP) is mediated by importin α, which recognizes its nuclear localization sequence (NLS), and importin β, which interacts with α and is in charge of the translocation of the NP/α/β complex through the nuclear pore. Herein, we characterize the assembly of a functional transport complex formed by full-length NP with importin α/β. Isothermal titration calorimetry (ITC) was used to analyze the thermodynamics of the interactions of importin α with β, α with NP, and the α/β heterodimer with NP. Our data show that binding of both importin α and α/β to NP is governed by a favorable enthalpic contribution and that NP can accommodate up to five importin molecules per NP pentamer. Phosphomimicking mutations of NP, which render the protein active in histone chaperoning, do not modulate the interaction with importin. Using small-angle X-ray scattering, we model the α/β heterodimer, NP/α, and NP/α/β solution structures, which reveal a glimpse of a complete nuclear import complex with an oligomeric cargo protein. The set of alternative models, equally well fitting the scattering data, yields asymmetric elongated particles that might represent consecutive geometries the complex can adopt when stepping through the nuclear pore.

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 #107: ; タイプ: dummy / ソフトウェア: Dammif / ダミー原子の半径: 6.80 A / カイ2乗値: 1.252161; 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 #108: ; タイプ: atomic / ソフトウェア: Sasref / カイ2乗値: 1.4161; 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: ImportinA_ImportinB / Sample MW: 160 kDa / 試料濃度: 0.50-1.00
• バッファ: 名称: 50 mM Tris-HCL / pH: 7.5 / 組成: 150 mM NaCl, 1.0 mM DTT

要素 #85

名称: ImpA_ImpB / タイプ: protein / 記述: ImportinA_ImportinB / 分子量: 160 / 分子数: 1 / 由来: Escherichia coli
配列:

VQVPPLSLEE IVQGMNSGDP ENELRCTQAA RKMLSRERNP PLNDIIEAGL IPKLVEFLSR HDNSTLQFEA AWALNIASGT SDQTKSVVDG GAIPAFISLI SSPHLHISEA VWALGNIAGD GPLYRDALIN CNVIPPLLAL VNPQTPLGYL NITWMLSNLC RNKNPYPPMS AVLQILPVLT QLMHHDDKDI LSDTCAMSYL TDGSNDRIDV VVKTGIVDRL IQLMYSPELS IVTPSLRTVG NIVTGTDKQT QAAIDAGVLS VLPQLLRHQK PSIQKEAAWA ISNIAAGPAP QIQQMITCGL LSPLVDLLNK GDFKAQKEAV WAVTNYTSGG TVEQVVQLVQ CGVLEPLLNL LTIKDSKTIL VILDAISNIF LAAEKLGEQE KLCLLVEELG GLEKIEALQT HDNHMVYHAA LALIEKYFME LITILEKTVS PDRLELEAAQ KFLERAAVEN LPTFLVELSR VLANPGNSQV ARVAAGLQIK NSLTSKDPDI KAQYQQRWLA IDANARREVK NYVLHTLGTE TYRPSSASQC VAGIACAEIP VNQWPELIPQ LVANVTNPNS TEHMKESTLE AIGYICQDID PEQLQDKSNE ILTAIIQGMR KEEPSNNVKL AATNALLNSL EFTKANFDKE SERHFIMQVV CEATQCPDTR VRVAALQNLV KIMSLYYQYM ETYMGPALFA ITIEAMKSDI DEVALQGIEF WSNVCDEEMD LAIEASEAAE QGRPPEHTSK FYAKGALQYL VPILTQTLTK QDENDDDDDW NPCKAAGVCL MLLATCCEDD IVPHVLPFIK EHIKNPDWRY RDAAVMAFGC ILEGPEPSQL KPLVIQAMPT LIELMKDPSV VVRDTAAWTV GRICELLPEA AINDVYLAPL LQCLIEGLSA EPRVASNVCW AFSSLAEAAY EAADVADDQE EPATYCLSSS FELIVQKLLE TTDRPDGHQN NLRSSAYESL MEIVKNSAKD CYPAVQKTTL VIMERLQQVL QMESHIQSTS DRIQFNDLQS LLCATLQNVL RKVQHQDALQ ISDVVMASLL RMFQSTAGSG GVQEDALMAV STLVEVLGGE FLKYMEAFKP FLGIGLKNYA EYQVCLAAVG LVGDLCRALQ SNIIPFCDEV MQLLLENLGN ENVHRSVKPQ ILSVFGDIAL AIGGEFKKYL EVVLNTLQQA SQAQVDKSDY DMVDYLNELR ESCLEAYTGI VQGLKGDQEN VHPDVMLVQP RVEFILSFID HIAGDEDHTD GVVACAAGLI GDLCTAFGKD VLKLVEARPM IHELLTEGRR SKTNKAKTLA RWATKELRKL KNQAAARLHR FKNKGKDSTE MRRRRIEVNV ELRKAKKDDQ MLKRRNVSSQ MANKEPSLIL EEPL

実験情報

• ビーム: 設備名称: DORIS III X33 / 地域: Hamburg / 国: Germany / 線源: X-ray synchrotron
• 検出器: 名称: Pilatus 2M

スキャン

タイトル: ImpA-ImpB / 測定日: 2009年10月29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	0.0775	6.237

距離分布関数 P(R)

ソフトウェア P(R): GNOM 4.5a / ポイント数: 320 /

	Min	Max
Q	0.1375	1.839
P(R) point	23	342
R	0	19

結果

カーブのタイプ: single_conc /

	実験値	Porod
分子量	185 kDa	-
体積	-	390 nm3

	P(R)	Guinier
前方散乱 I0	81	80
慣性半径, Rg	5.7 nm	5.65 nm

	Min	Max
D	-	19
Guinier point	12	67