SASDAD5: Nucleoplasmin_ImpA_ImpB (Nucleoplasmin_importinA_importinB, NP_Im...

Basic information

• Entry: Database: SASBDB / ID: SASDAD5

Sample

Nucleoplasmin_ImpA_ImpB

• Nucleoplasmin_importinA_importinB (protein), NP_ImpA_ImpB, Escherichia coli

• Biological species: Escherichia coli (E. coli)
• Citation: Journal: Biochemistry / Year: 2010; Title: Recognition of nucleoplasmin by its nuclear transport receptor importin α/β: insights into a complete import complex.; Authors: Jorge Falces / Igor Arregi / Petr V Konarev / María A Urbaneja / Dmitri I Svergun / Stefka G Taneva / Sonia Bañuelos / ; Abstract: 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.

Contact author

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

Models

• Model #109: ; Type: dummy / Software: Dammif / Radius of dummy atoms: 5.00 A / Chi-square value: 1.540081; Search similar-shape structures of this assembly by Omokage search (details)
• Model #110: ; Type: atomic / Software: Sasref / Chi-square value: 3.8809; Search similar-shape structures of this assembly by Omokage search (details)

Sample

• Sample: Name: Nucleoplasmin_ImpA_ImpB / Sample MW: 900 kDa / Specimen concentration: 1.00-4.00
• Buffer: Name: 50 mM Tris-HCL / pH: 7.5 / Composition: 150 mM NaCl, 1.0 mM DTT

Entity #86

Name: NP_ImpA_ImpB / Type: protein / Description: Nucleoplasmin_importinA_importinB / Formula weight: 900 / Source: Escherichia coli
Sequence:

MASTVSNTSK LEKPVVSLIW GCELNEQNKT FEFKVEDDEE KCEHQLALRT VCLGDKAKDE FHIVEIVTQE EGAEKSVPIA TLKPSILPMA TMVGIELTPP VTFRLKAGSG PLYISGQHVA MASTVSNTSK LEKPVMASTV SNTSKLEKPV MASTVSNTSK LEKPVMASTV SNTSKLEKPV VSLIWGCELN EQNKTFEFKV EDDEEKCEHQ LALRTVCLGD KAKDEFHIVE IVTQEEGAEK SVPIATLKPS ILPMATMVGI ELTPPVTFRL KAGSGPLYIS GQHVAVSLIW GCELNEQNKT FEFKVEDDEE KCEHQLALRT VCLGDKAKDE FHIVEIVTQE EGAEKSVPIA TLKPSILPMA TMVGIELTPP VTFRLKAGSG PLYISGQHVA VSLIWGCELN EQNKTFEFKV EDDEEKCEHQ LALRTVCLGD KAKDEFHIVE IVTQEEGAEK SVPIATLKPS ILPMATMVGI ELTPPVTFRL KAGSGPLYIS GQHVAVSLIW GCELNEQNKT FEFKVEDDEE KCEHQLALRT VCLGDKAKDE FHIVEIVTQE EGAEKSVPIA TLKPSILPMA TMVGIELTPP VTFRLKAGSG PLYISGQHVA MEEDYSWAEE EDEGEAEGEE EEEEEEDQES PPKAVKRPAA TKKAGQAKKK KLDKEDESSE EDSPTKKGKG AGRGRKPAAK KMEEDYSWAE EEDEGEAEGE EEEEEEEDQE SPPKAVKRPA ATKKAGQAKK KKLDKEDESS EEDSPTKKGK GAGRGRKPAA KKMEEDYSWA EEEDEGEAEG EEEEEEEEDQ ESPPKAVKRP AATKKAGQAK KKKLDKEDES SEEDSPTKKG KGAGRGRKPA AKKMEEDYSW AEEEDEGEAE GEEEEEEEED QESPPKAVKR PAATKKAGQA KKKKLDKEDE SSEEDSPTKK GKGAGRGRKP AAKKMEEDYS WAEEEDEGEA EGEEEEEEEE DQESPPKAVK RPAATKKAGQ AKKKKLDKED ESSEEDSPTK KGKGAGRGRK PAAKKVQVPP LSLEEIVQGM NSGDPENELR CTQAARKMLS RERNPPLNDI IEAGLIPKLV EFLSRHDNST LQFEAAWALN IASGTSDQTK SVVDGGAIPA FISLISSPHL HISEAVWALG NIAGDGPLYR DALINCNVIP PLLALVNPQT PLGYLNITWM LSNLCRNKNP YPPMSAVLQI LPVLTQLMHH DDKDILSDTC AMSYLTDGSN DRIDVVVKTG IVDRLIQLMY SPELSIVTPS LRTVGNIVTG TDKQTQAAID AGVLSVLPQL LRHQKPSIQK EAAWAISNIA AGPAPQIQQM ITCGLLSPLV DLLNKGDFKA QKEAVWAVTN YTSGGTVEQV VQLVQCGVLE PLLNLLTIKD SKTILVILDA ISNIFLAAEK LGEQEKLCLL VEELGGLEKI EALQTHDNHM VYHAALALIE KYFVQVPPLS LEEIVQGMNS GDPENELRCT QAARKMLSRE RNPPLNDIIE AGLIPKLVEF LSRHDNSTLQ FEAAWALNIA SGTSDQTKSV VDGGAIPAFI SLISSPHLHI SEAVWALGNI AGDGPLYRDA LINCNVIPPL LALVNPQTPL GYLNITWMLS NLCRNKNPYP PMSAVLQILP VLTQLMHHDD KDILSDTCAM SYLTDGSNDR IDVVVKTGIV DRLIQLMYSP ELSIVTPSLR TVGNIVTGTD KQTQAAIDAG VLSVLPQLLR HQKPSIQKEA AWAISNIAAG PAPQIQQMIT CGLLSPLVDL LNKGDFKAQK EAVWAVTNYT SGGTVEQVVQ LVQCGVLEPL LNLLTIKDSK TILVILDAIS NIFLAAEKLG EQEKLCLLVE ELGGLEKIEA LQTHDNHMVY HAALALIEKY FVQVPPLSLE EIVQGMNSGD PENELRCTQA ARKMLSRERN PPLNDIIEAG LIPKLVEFLS RHDNSTLQFE AAWALNIASG TSDQTKSVVD GGAIPAFISL ISSPHLHISE AVWALGNIAG DGPLYRDALI NCNVIPPLLA LVNPQTPLGY LNITWMLSNL CRNKNPYPPM SAVLQILPVL TQLMHHDDKD ILSDTCAMSY LTDGSNDRID VVVKTGIVDR LIQLMYSPEL SIVTPSLRTV GNIVTGTDKQ TQAAIDAGVL SVLPQLLRHQ KPSIQKEAAW AISNIAAGPA PQIQQMITCG LLSPLVDLLN KGDFKAQKEA VWAVTNYTSG GTVEQVVQLV QCGVLEPLLN LLTIKDSKTI LVILDAISNI FLAAEKLGEQ EKLCLLVEEL GGLEKIEALQ THDNHMVYHA ALALIEKYFV QVPPLSLEEI VQGMNSGDPE NELRCTQAAR KMLSRERNPP LNDIIEAGLI PKLVEFLSRH DNSTLQFEAA WALNIASGTS DQTKSVVDGG AIPAFISLIS SPHLHISEAV WALGNIAGDG PLYRDALINC NVIPPLLALV NPQTPLGYLN ITWMLSNLCR NKNPYPPMSA VLQILPVLTQ LMHHDDKDIL SDTCAMSYLT DGSNDRIDVV VKTGIVDRLI QLMYSPELSI VTPSLRTVGN IVTGTDKQTQ AAIDAGVLSV LPQLLRHQKP SIQKEAAWAI SNIAAGPAPQ IQQMITCGLL SPLVDLLNKG DFKAQKEAVW AVTNYTSGGT VEQVVQLVQC GVLEPLLNLL TIKDSKTILV ILDAISNIFL AAEKLGEQEK LCLLVEELGG LEKIEALQTH DNHMVYHAAL ALIEKYFVQV PPLSLEEIVQ GMNSGDPENE LRCTQAARKM LSRERNPPLN DIIEAGLIPK LVEFLSRHDN STLQFEAAWA LNIASGTSDQ TKSVVDGGAI PAFISLISSP HLHISEAVWA LGNIAGDGPL YRDALINCNV IPPLLALVNP QTPLGYLNIT WMLSNLCRNK NPYPPMSAVL QILPVLTQLM HHDDKDILSD TCAMSYLTDG SNDRIDVVVK TGIVDRLIQL MYSPELSIVT PSLRTVGNIV TGTDKQTQAA IDAGVLSVLP QLLRHQKPSI QKEAAWAISN IAAGPAPQIQ QMITCGLLSP LVDLLNKGDF KAQKEAVWAV TNYTSGGTVE QVVQLVQCGV LEPLLNLLTI KDSKTILVIL DAISNIFLAA EKLGEQEKLC LLVEELGGLE KIEALQTHDN HMVYHAALAL IEKYFMELIT ILEKTVSPDR LELEAAQKFL ERAAVENLPT FLVELSRVLA NPGNSQVARV AAGLQIKNSL TSKDPDIKAQ YQQRWLAIDA NARREVKNYV LHTLGTETYR PSSASQCVAG IACAEIPVNQ WPELIPQLVA NVTNPNSTEH MKESTLEAIG YICQDIDPEQ LQDKSNEILT AIIQGMRKEE PSNNVKLAAT NALLNSLEFT KANFDKESER HFIMQVVCEA TQCPDTRVRV AALQNLVKIM SLYYQYMETY MGPALFAITI EAMKSDIDEV ALQGIEFWSN VCDEEMDLAI EASEAAEQGR PPEHTSKFYA KGALQYLVPI LTQTLTKQDE NDDDDDWNPC KAAGVCLMLL ATCCEDDIVP HVLPFIKEHI KNPDWRYRDA AVMAFGCILE GPEPSQLKPL VIQAMPTLIE LMKDPSVVVR DTAAWTVGRI CELLPEAAIN DVYLAPLLQC LIEGLSAEPR VASNVCWAFS SLAEAAYEAA DVADDQEEPA TYCLSSSFEL IVQKLLETTD RPDGHQNNLR SSAYESLMEI VKNSAKDCYP AVQKTTLVIM ERLQQVLQME SHIQSTSDRI QFNDLQSLLC ATLQNVLRKV QHQDALQISD VVMASLLRMF QSTAGSGGVQ EDALMAVSTL VEVLGGEFLK YMEAFKPFLG IGLKNYAEYQ VCLAAVGLVG DLCRALQSNI IPFCDEVMQL LLENLGNENV HRSVKPQILS VFGDIALAIG GEFKKYLEVV LNTLQQASQA QVDKSDYDMV DYLNELRESC LEAYTGIVQG LKGDQENVHP DVMLVQPRVE FILSFIDHIA GDEDHTDGVV ACAAGLIGDL CTAFGKDVLK LVEARPMIHE LLTEGRRSKT NKAKTLARWA TKELRKLKNQ AAARLHRFKN KGKDSTEMRR RRIEVNVELR KAKKDDQMLK RRNVSSQMAN KEPSLILEEP LMELITILEK TVSPDRLELE AAQKFLERAA VENLPTFLVE LSRVLANPGN SQVARVAAGL QIKNSLTSKD PDIKAQYQQR WLAIDANARR EVKNYVLHTL GTETYRPSSA SQCVAGIACA EIPVNQWPEL IPQLVANVTN PNSTEHMKES TLEAIGYICQ DIDPEQLQDK SNEILTAIIQ GMRKEEPSNN VKLAATNALL NSLEFTKANF DKESERHFIM QVVCEATQCP DTRVRVAALQ NLVKIMSLYY QYMETYMGPA LFAITIEAMK SDIDEVALQG IEFWSNVCDE EMDLAIEASE AAEQGRPPEH TSKFYAKGAL QYLVPILTQT LTKQDENDDD DDWNPCKAAG VCLMLLATCC EDDIVPHVLP FIKEHIKNPD WRYRDAAVMA FGCILEGPEP SQLKPLVIQA MPTLIELMKD PSVVVRDTAA WTVGRICELL PEAAINDVYL APLLQCLIEG LSAEPRVASN VCWAFSSLAE AAYEAADVAD DQEEPATYCL SSSFELIVQK LLETTDRPDG HQNNLRSSAY ESLMEIVKNS AKDCYPAVQK TTLVIMERLQ QVLQMESHIQ STSDRIQFND LQSLLCATLQ NVLRKVQHQD ALQISDVVMA SLLRMFQSTA GSGGVQEDAL MAVSTLVEVL GGEFLKYMEA FKPFLGIGLK NYAEYQVCLA AVGLVGDLCR ALQSNIIPFC DEVMQLLLEN LGNENVHRSV KPQILSVFGD IALAIGGEFK KYLEVVLNTL QQASQAQVDK SDYDMVDYLN ELRESCLEAY TGIVQGLKGD QENVHPDVML VQPRVEFILS FIDHIAGDED HTDGVVACAA GLIGDLCTAF GKDVLKLVEA RPMIHELLTE GRRSKTNKAK TLARWATKEL RKLKNQAAAR LHRFKNKGKD STEMRRRRIE VNVELRKAKK DDQMLKRRNV SSQMANKEPS LILEEPLMEL ITILEKTVSP DRLELEAAQK FLERAAVENL PTFLVELSRV LANPGNSQVA RVAAGLQIKN SLTSKDPDIK AQYQQRWLAI DANARREVKN YVLHTLGTET YRPSSASQCV AGIACAEIPV NQWPELIPQL VANVTNPNST EHMKESTLEA IGYICQDIDP EQLQDKSNEI LTAIIQGMRK EEPSNNVKLA ATNALLNSLE FTKANFDKES ERHFIMQVVC EATQCPDTRV RVAALQNLVK IMSLYYQYME TYMGPALFAI TIEAMKSDID EVALQGIEFW SNVCDEEMDL AIEASEAAEQ GRPPEHTSKF YAKGALQYLV PILTQTLTKQ DENDDDDDWN PCKAAGVCLM LLATCCEDDI VPHVLPFIKE HIKNPDWRYR DAAVMAFGCI LEGPEPSQLK PLVIQAMPTL IELMKDPSVV VRDTAAWTVG RICELLPEAA INDVYLAPLL QCLIEGLSAE PRVASNVCWA FSSLAEAAYE AADVADDQEE PATYCLSSSF ELIVQKLLET TDRPDGHQNN LRSSAYESLM EIVKNSAKDC YPAVQKTTLV IMERLQQVLQ MESHIQSTSD RIQFNDLQSL LCATLQNVLR KVQHQDALQI SDVVMASLLR MFQSTAGSGG VQEDALMAVS TLVEVLGGEF LKYMEAFKPF LGIGLKNYAE YQVCLAAVGL VGDLCRALQS NIIPFCDEVM QLLLENLGNE NVHRSVKPQI LSVFGDIALA IGGEFKKYLE VVLNTLQQAS QAQVDKSDYD MVDYLNELRE SCLEAYTGIV QGLKGDQENV HPDVMLVQPR VEFILSFIDH IAGDEDHTDG VVACAAGLIG DLCTAFGKDV LKLVEARPMI HELLTEGRRS KTNKAKTLAR WATKELRKLK NQAAARLHRF KNKGKDSTEM RRRRIEVNVE LRKAKKDDQM LKRRNVSSQM ANKEPSLILE EPLMELITIL EKTVSPDRLE LEAAQKFLER AAVENLPTFL VELSRVLANP GNSQVARVAA GLQIKNSLTS KDPDIKAQYQ QRWLAIDANA RREVKNYVLH TLGTETYRPS SASQCVAGIA CAEIPVNQWP ELIPQLVANV TNPNSTEHMK ESTLEAIGYI CQDIDPEQLQ DKSNEILTAI IQGMRKEEPS NNVKLAATNA LLNSLEFTKA NFDKESERHF IMQVVCEATQ CPDTRVRVAA LQNLVKIMSL YYQYMETYMG PALFAITIEA MKSDIDEVAL QGIEFWSNVC DEEMDLAIEA SEAAEQGRPP EHTSKFYAKG ALQYLVPILT QTLTKQDEND DDDDWNPCKA AGVCLMLLAT CCEDDIVPHV LPFIKEHIKN PDWRYRDAAV MAFGCILEGP EPSQLKPLVI QAMPTLIELM KDPSVVVRDT AAWTVGRICE LLPEAAINDV YLAPLLQCLI EGLSAEPRVA SNVCWAFSSL AEAAYEAADV ADDQEEPATY CLSSSFELIV QKLLETTDRP DGHQNNLRSS AYESLMEIVK NSAKDCYPAV QKTTLVIMER LQQVLQMESH IQSTSDRIQF NDLQSLLCAT LQNVLRKVQH QDALQISDVV MASLLRMFQS TAGSGGVQED ALMAVSTLVE VLGGEFLKYM EAFKPFLGIG LKNYAEYQVC LAAVGLVGDL CRALQSNIIP FCDEVMQLLL ENLGNENVHR SVKPQILSVF GDIALAIGGE FKKYLEVVLN TLQQASQAQV DKSDYDMVDY LNELRESCLE AYTGIVQGLK GDQENVHPDV MLVQPRVEFI LSFIDHIAGD EDHTDGVVAC AAGLIGDLCT AFGKDVLKLV EARPMIHELL TEGRRSKTNK AKTLARWATK ELRKLKNQAA ARLHRFKNKG KDSTEMRRRR IEVNVELRKA KKDDQMLKRR NVSSQMANKE PSLILEEPLM ELITILEKTV SPDRLELEAA QKFLERAAVE NLPTFLVELS RVLANPGNSQ VARVAAGLQI KNSLTSKDPD IKAQYQQRWL AIDANARREV KNYVLHTLGT ETYRPSSASQ CVAGIACAEI PVNQWPELIP QLVANVTNPN STEHMKESTL EAIGYICQDI DPEQLQDKSN EILTAIIQGM RKEEPSNNVK LAATNALLNS LEFTKANFDK ESERHFIMQV VCEATQCPDT RVRVAALQNL VKIMSLYYQY METYMGPALF AITIEAMKSD IDEVALQGIE FWSNVCDEEM DLAIEASEAA EQGRPPEHTS KFYAKGALQY LVPILTQTLT KQDENDDDDD WNPCKAAGVC LMLLATCCED DIVPHVLPFI KEHIKNPDWR YRDAAVMAFG CILEGPEPSQ LKPLVIQAMP TLIELMKDPS VVVRDTAAWT VGRICELLPE AAINDVYLAP LLQCLIEGLS AEPRVASNVC WAFSSLAEAA YEAADVADDQ EEPATYCLSS SFELIVQKLL ETTDRPDGHQ NNLRSSAYES LMEIVKNSAK DCYPAVQKTT LVIMERLQQV LQMESHIQST SDRIQFNDLQ SLLCATLQNV LRKVQHQDAL QISDVVMASL LRMFQSTAGS GGVQEDALMA VSTLVEVLGG EFLKYMEAFK PFLGIGLKNY AEYQVCLAAV GLVGDLCRAL QSNIIPFCDE VMQLLLENLG NENVHRSVKP QILSVFGDIA LAIGGEFKKY LEVVLNTLQQ ASQAQVDKSD YDMVDYLNEL RESCLEAYTG IVQGLKGDQE NVHPDVMLVQ PRVEFILSFI DHIAGDEDHT DGVVACAAGL IGDLCTAFGK DVLKLVEARP MIHELLTEGR RSKTNKAKTL ARWATKELRK LKNQAAARLH RFKNKGKDST EMRRRRIEVN VELRKAKKDD QMLKRRNVSS QMANKEPSLI LEEPL

Experimental information

• Beam: Instrument name: DORIS III X33 / City: Hamburg / 国: Germany / Type of source: X-ray synchrotron
• Detector: Name: Pilatus 2M

Scan

Title: NP_ImpA_impB / Measurement date: May 23, 2008 / Storage temperature: 10 °C / Cell temperature: 10 °C / Exposure time: 120 sec. / Number of frames: 2 / Unit: 1/nm /

	Min	Max
Q	0.107	1.4674

Distance distribution function P(R)

Sofotware P(R): GNOM 4.5a / Number of points: 245 /

	Min	Max
Q	0.107	1.092
P(R) point	1	245
R	0	25

Result

D max: 28 / Type of curve: single_conc /

	Experimental	Porod
MW	860 kDa	-
Volume	-	1800 nm3

	P(R)	Guinier
Forward scattering, I0	4072	4070
Radius of gyration, Rg	8.57 nm	8.55 nm