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- SASDCR9: Kif2A-tubulin-DARPin complex in presence of AMP-PNP -

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Entry
Database: SASBDB / ID: SASDCR9
SampleKif2A-tubulin-DARPin complex in presence of AMP-PNP
  • Designed Ankyrin Repeat Protein D1 (protein), DARPin D1, synthetic construct
  • Kinesin-like protein KIF2A (protein), Kif2A, Homo sapiens
  • Tubulin alpha-1B chain (protein), alpha tubulin, Bos taurus
  • Tubulin beta-2B chain (protein), beta tubulin, Bos taurus
Biological speciessynthetic construct (others)
Homo sapiens (human)
Bos taurus (cattle)

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Models

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Sample

SampleName: Kif2A-tubulin-DARPin complex in presence of AMP-PNP / Specimen concentration: 3.7 mg/ml / Entity id: 875 / 890 / 902 / 903
BufferName: HEPES 20 mM, MgCl2 1mM, NaCl 150mM / pH: 7.2
Entity #875Name: DARPin D1 / Type: protein / Description: Designed Ankyrin Repeat Protein D1 / Formula weight: 17.886 / Num. of mol.: 1 / Source: synthetic construct
Sequence:
MGSHHHHHHG SDLGKKLLEA ARAGQDDEVR ILMANGADVN ATDASGLTPL HLAATYGHLE IVEVLLKHGA DVNAIDIMGS TPLHLAALIG HLEIVEVLLK HGADVNAVDT WGDTPLHLAA IMGHLEIVEV LLKHGADVNA QDKFGKTAFD ISIDNGNEDL AEILQKLN
Entity #890Name: Kif2A / Type: protein / Description: Kinesin-like protein KIF2A / Formula weight: 47.834 / Num. of mol.: 1 / Source: Homo sapiens
Sequence: MGSSHHHHHH SSGLVPRGSS RRKSNCVKEV EKLQEKREKR RLQQQELREK RAQDVDATNP NYEIMCMIRD FRGSLDYRPL TTADPIDEHR ICVCVRKRPL NKKETQMKDL DVITIPSKDV VMVHEPKQKV DLTRYLENQT FRFDYAFDDS APNEMVYRFT ARPLVETIFE ...Sequence:
MGSSHHHHHH SSGLVPRGSS RRKSNCVKEV EKLQEKREKR RLQQQELREK RAQDVDATNP NYEIMCMIRD FRGSLDYRPL TTADPIDEHR ICVCVRKRPL NKKETQMKDL DVITIPSKDV VMVHEPKQKV DLTRYLENQT FRFDYAFDDS APNEMVYRFT ARPLVETIFE RGMATCFAYG QTGSGKTHTM GGDFSGKNQD CSKGIYALAA RDVFLMLKKP NYKKLELQVY ATFFEIYSGK VFDLLNRKTK LRVLEDGKQQ VQVVGLQERE VKCVEDVLKL IDIGNSCRTS GQTSANAHSS RSHAVFQIIL RRKGKLHGKF SLIDLAGNER GADTSSADRQ TRLEGAEINK SLLALKECIR ALGRNKPHTP FRASKLTQVL RDSFIGENSR TCMIATISPG MASCENTLNT LRYANRVKEL
Entity #902Name: alpha tubulin / Type: protein / Description: Tubulin alpha-1B chain / Formula weight: 50.151 / Num. of mol.: 2 / Source: Bos taurus
Sequence: MRECISIHVG QAGVQIGNAC WELYCLEHGI QPDGQMPSDK TIGGGDDSFN TFFSETGAGK HVPRAVFVDL EPTVIDEVRT GTYRQLFHPE QLITGKEDAA NNYARGHYTI GKEIIDLVLD RIRKLADQCT GLQGFLVFHS FGGGTGSGFT SLLMERLSVD YGKKSKLEFS ...Sequence:
MRECISIHVG QAGVQIGNAC WELYCLEHGI QPDGQMPSDK TIGGGDDSFN TFFSETGAGK HVPRAVFVDL EPTVIDEVRT GTYRQLFHPE QLITGKEDAA NNYARGHYTI GKEIIDLVLD RIRKLADQCT GLQGFLVFHS FGGGTGSGFT SLLMERLSVD YGKKSKLEFS IYPAPQVSTA VVEPYNSILT THTTLEHSDC AFMVDNEAIY DICRRNLDIE RPTYTNLNRL ISQIVSSITA SLRFDGALNV DLTEFQTNLV PYPRIHFPLA TYAPVISAEK AYHEQLSVAE ITNACFEPAN QMVKCDPRHG KYMACCLLYR GDVVPKDVNA AIATIKTKRS IQFVDWCPTG FKVGINYQPP TVVPGGDLAK VQRAVCMLSN TTAIAEAWAR LDHKFDLMYA KRAFVHWYVG EGMEEGEFSE AREDMAALEK DYEEVGVDSV EGEGEEEGEE Y
Entity #903Name: beta tubulin / Type: protein / Description: Tubulin beta-2B chain / Formula weight: 49.936 / Num. of mol.: 2 / Source: Bos taurus
Sequence: MREIVHIQAG QCGNQIGAKF WEVISDEHGI DPTGSYHGDS DLQLERINVY YNEATGNKYV PRAILVDLEP GTMDSVRSGP FGQIFRPDNF VFGQSGAGNN WAKGHYTEGA ELVDSVLDVV RKESESCDCL QGFQLTHSLG GGTGSGMGTL LISKIREEYP DRIMNTFSVM ...Sequence:
MREIVHIQAG QCGNQIGAKF WEVISDEHGI DPTGSYHGDS DLQLERINVY YNEATGNKYV PRAILVDLEP GTMDSVRSGP FGQIFRPDNF VFGQSGAGNN WAKGHYTEGA ELVDSVLDVV RKESESCDCL QGFQLTHSLG GGTGSGMGTL LISKIREEYP DRIMNTFSVM PSPKVSDTVV EPYNATLSVH QLVENTDETY SIDNEALYDI CFRTLKLTTP TYGDLNHLVS ATMSGVTTCL RFPGQLNADL RKLAVNMVPF PRLHFFMPGF APLTSRGSQQ YRALTVPELT QQMFDSKNMM AACDPRHGRY LTVAAIFRGR MSMKEVDEQM LNVQNKNSSY FVEWIPNNVK TAVCDIPPRG LKMSATFIGN STAIQELFKR ISEQFTAMFR RKAFLHWYTG EGMDEMEFTE AESNMNDLVS EYQQYQDATA DEQGEFEEEE GEDEA

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Experimental information

BeamInstrument name: Cornell High Energy Synchrotron Source (CHESS) G1
City: Ithaca, NY / 国: USA / Type of source: X-ray synchrotron / Wavelength: 0.1267 Å / Dist. spec. to detc.: 1.47 mm
DetectorName: Finger Lakes CCD / Type: CCD
ScanTitle: Kif2A-tubulin-DARPin complex in presence of AMP-PNP / Measurement date: May 16, 2016 / Storage temperature: 4 °C / Cell temperature: 22 °C / Exposure time: 2 sec. / Number of frames: 1200 / Unit: 1/A /
MinMax
Q0.0086 0.2735
ResultType of curve: sec
Comments: For the SEC-SAXS analysis, a mixture of Kif2A, tubulin and DARPin was passed continuously through an X-ray sample cell via an in-line size exclusion column (GE Superdex 200 10/300 GL) at a ...Comments: For the SEC-SAXS analysis, a mixture of Kif2A, tubulin and DARPin was passed continuously through an X-ray sample cell via an in-line size exclusion column (GE Superdex 200 10/300 GL) at a flow rate of 1 ml/min. The column was pre-equilibrated with running buffer consisting of 20 mM HEPES, 150 mM NaCl, 1 mM MgCl2 pH 7.2. The protein sample was prepared at 25 μM tubulin, 27 μM DARP in and 20 μM Kif2A and injected into a 100 μL loop. Approximately 1200 two-second exposures were collected per sample, and 100 buffer profiles preceding the elution peaks were averaged and used for background subtraction.
ExperimentalPorod
MW258.5 kDa279.7 kDa
Volume-374 nm3

P(R)P(R) errorGuinierGuinier error
Forward scattering, I00.0298 0.00029 0.02949 0.0002
Radius of gyration, Rg5.6 nm0.03 5.41 nm0.05

MinMax
D-19.5
Guinier point4 26

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