+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-25387 | |||||||||
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Title | Structure of a Protein Complex in Cell Division | |||||||||
Map data | Structure of a Protein Complex in Cell Division | |||||||||
Sample |
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Keywords | Cell Division / Mitosis / Microtubules / CELL CYCLE | |||||||||
Function / homology | Function and homology information HAUS complex / regulation of microtubule nucleation / microtubule minus-end binding / nuclear microtubule / mitotic spindle microtubule / microtubule nucleation / thioesterase binding / centrosome cycle / spindle assembly / Loss of Nlp from mitotic centrosomes ...HAUS complex / regulation of microtubule nucleation / microtubule minus-end binding / nuclear microtubule / mitotic spindle microtubule / microtubule nucleation / thioesterase binding / centrosome cycle / spindle assembly / Loss of Nlp from mitotic centrosomes / Loss of proteins required for interphase microtubule organization from the centrosome / Recruitment of mitotic centrosome proteins and complexes / Recruitment of NuMA to mitotic centrosomes / Anchoring of the basal body to the plasma membrane / AURKA Activation by TPX2 / mitotic spindle / spindle pole / microtubule cytoskeleton organization / Regulation of PLK1 Activity at G2/M Transition / microtubule binding / cell division / centrosome / nucleolus / plasma membrane / cytosol / cytoplasm Similarity search - Function | |||||||||
Biological species | Homo sapiens (human) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 8.0 Å | |||||||||
Authors | Gabel CA / Chang L | |||||||||
Funding support | 1 items
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Citation | Journal: Nat Commun / Year: 2022 Title: Molecular architecture of the augmin complex. Authors: Clinton A Gabel / Zhuang Li / Andrew G DeMarco / Ziguo Zhang / Jing Yang / Mark C Hall / David Barford / Leifu Chang / Abstract: Accurate segregation of chromosomes during mitosis depends on the correct assembly of the mitotic spindle, a bipolar structure composed mainly of microtubules. The augmin complex, or homologous to ...Accurate segregation of chromosomes during mitosis depends on the correct assembly of the mitotic spindle, a bipolar structure composed mainly of microtubules. The augmin complex, or homologous to augmin subunits (HAUS) complex, is an eight-subunit protein complex required for building robust mitotic spindles in metazoa. Augmin increases microtubule density within the spindle by recruiting the γ-tubulin ring complex (γ-TuRC) to pre-existing microtubules and nucleating branching microtubules. Here, we elucidate the molecular architecture of augmin by single particle cryo-electron microscopy (cryo-EM), computational methods, and crosslinking mass spectrometry (CLMS). Augmin's highly flexible structure contains a V-shaped head and a filamentous tail, with the head existing in either extended or contracted conformational states. Our work highlights how cryo-EM, complemented by computational advances and CLMS, can elucidate the structure of a challenging protein complex and provides insights into the function of augmin in mediating microtubule branching nucleation. | |||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_25387.map.gz | 215.8 MB | EMDB map data format | |
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Header (meta data) | emd-25387-v30.xml emd-25387.xml | 22.6 KB 22.6 KB | Display Display | EMDB header |
Images | emd_25387.png | 35.1 KB | ||
Filedesc metadata | emd-25387.cif.gz | 8 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-25387 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-25387 | HTTPS FTP |
-Validation report
Summary document | emd_25387_validation.pdf.gz | 311.5 KB | Display | EMDB validaton report |
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Full document | emd_25387_full_validation.pdf.gz | 311.1 KB | Display | |
Data in XML | emd_25387_validation.xml.gz | 7.5 KB | Display | |
Data in CIF | emd_25387_validation.cif.gz | 8.7 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-25387 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-25387 | HTTPS FTP |
-Related structure data
Related structure data | 7sqkMC M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_25387.map.gz / Format: CCP4 / Size: 421.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Annotation | Structure of a Protein Complex in Cell Division | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.384 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Sample components
-Entire : Holocomplex of the Homologous to Augment Subunits Complex
Entire | Name: Holocomplex of the Homologous to Augment Subunits Complex |
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Components |
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-Supramolecule #1: Holocomplex of the Homologous to Augment Subunits Complex
Supramolecule | Name: Holocomplex of the Homologous to Augment Subunits Complex type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 378 kDa/nm |
-Macromolecule #1: HAUS augmin-like complex subunit 1
Macromolecule | Name: HAUS augmin-like complex subunit 1 / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 31.910348 KDa |
Recombinant expression | Organism: Trichoplusia ni (cabbage looper) |
Sequence | String: MEPQEERETQ VAAWLKKIFG DHPIPQYEVN PRTTEILHHL SERNRVRDRD VYLVIEDLKQ KASEYESEAK YLQDLLMESV NFSPANLSS TGSRYLNALV DSAVALETKD TSLASFIPAV NDLTSDLFRT KSKSEEIKIE LEKLEKNLTA TLVLEKCLQE D VKKAELHL ...String: MEPQEERETQ VAAWLKKIFG DHPIPQYEVN PRTTEILHHL SERNRVRDRD VYLVIEDLKQ KASEYESEAK YLQDLLMESV NFSPANLSS TGSRYLNALV DSAVALETKD TSLASFIPAV NDLTSDLFRT KSKSEEIKIE LEKLEKNLTA TLVLEKCLQE D VKKAELHL STERAKVDNR RQNMDFLKAK SEEFRFGIKA AEEQLSARGM DASLSHQSLV ALSEKLARLK QQTIPLKKKL ES YLDLMPN PSLAQVKIEE AKRELDSIEA ELTRRVDMME L UniProtKB: HAUS augmin-like complex subunit 1 |
-Macromolecule #2: HAUS augmin-like complex subunit 2
Macromolecule | Name: HAUS augmin-like complex subunit 2 / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 32.487453 KDa |
Recombinant expression | Organism: Trichoplusia ni (cabbage looper) |
Sequence | String: MHHHHHHHHP QLAMWSHPQF EKGGGSGGGS GGGSWSHPQF EKENLYFQSM AAANPWDPAS APNGAGLVLG HFIASGMVNQ EMLNMSKKT VSCFVNFTRL QQITNIQAEI YQKNLEIELL KLEKDTADVV HPFFLAQKCH TLQSMNNHLE AVLKEKRSLR Q RLLKPMCQ ...String: MHHHHHHHHP QLAMWSHPQF EKGGGSGGGS GGGSWSHPQF EKENLYFQSM AAANPWDPAS APNGAGLVLG HFIASGMVNQ EMLNMSKKT VSCFVNFTRL QQITNIQAEI YQKNLEIELL KLEKDTADVV HPFFLAQKCH TLQSMNNHLE AVLKEKRSLR Q RLLKPMCQ ENLPIEAVYH RYMVHLLELA VTFIERLETH LETIRNIPHL AANLKKMNQA LAKMDILVTE TEELAENILK WR KQQNEVS SCIPKILAEE SYLYKHDIIM PPLPFTSKVH VQTINAK UniProtKB: HAUS augmin-like complex subunit 2 |
-Macromolecule #3: HAUS augmin-like complex subunit 3
Macromolecule | Name: HAUS augmin-like complex subunit 3 / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 69.74018 KDa |
Recombinant expression | Organism: Trichoplusia ni (cabbage looper) |
Sequence | String: MSCGNEFVET LKKIGYPKAD NLNGEDFDWL FEGVEDESFL KWFCGNVNEQ NVLSERELEA FSILQKSGKP ILEGAALDEA LKTCKTSDL KTPRLDDKEL EKLEDEVQTL LKLKNLKIQR RNKCQLMASV TSHKSLRLNA KEEEATKKLK QSQGILNAMI T KISNELQA ...String: MSCGNEFVET LKKIGYPKAD NLNGEDFDWL FEGVEDESFL KWFCGNVNEQ NVLSERELEA FSILQKSGKP ILEGAALDEA LKTCKTSDL KTPRLDDKEL EKLEDEVQTL LKLKNLKIQR RNKCQLMASV TSHKSLRLNA KEEEATKKLK QSQGILNAMI T KISNELQA LTDEVTQLMM FFRHSNLGQG TNPLVFLSQF SLEKYLSQEE QSTAALTLYT KKQFFQGIHE VVESSNEDNF QL LDIQTPS ICDNQEILEE RRLEMARLQL AYICAQHQLI HLKASNSSMK SSIKWAEESL HSLTSKAVDK ENLDAKISSL TSE IMKLEK EVTQIKDRSL PAVVRENAQL LNMPVVKGDF DLQIAKQDYY TARQELVLNQ LIKQKASFEL LQLSYEIELR KHRD IYRQL ENLVQELSQS NMMLYKQLEM LTDPSVSQQI NPRNTIDTKD YSTHRLYQVL EGENKKKELF LTHGNLEEVA EKLKQ NISL VQDQLAVSAQ EHSFFLSKRN KDVDMLCDTL YQGGNQLLLS DQELTEQFHK VESQLNKLNH LLTDILADVK TKRKTL ANN KLHQMEREFY VYFLKDEDYL KDIVENLETQ SKIKAVSLED UniProtKB: HAUS augmin-like complex subunit 3 |
-Macromolecule #4: Isoform 4 of HAUS augmin-like complex subunit 4
Macromolecule | Name: Isoform 4 of HAUS augmin-like complex subunit 4 / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 37.269805 KDa |
Recombinant expression | Organism: Trichoplusia ni (cabbage looper) |
Sequence | String: MASGDFCSPG EGMEILQQVC SKQLPPCNLS KEDLLQNPYF SKLLLNLSQH VDESGLSLTL AKEQAQAWKE VRLHKTTWLR SEILHRVIQ ELLVDYYVKI QDTNVTSEDK KDFVWMRARL QQEVEEQLKK KCFTLLCYYD PNSDADSETV KAAKVWKLAE V LVGEQQQC ...String: MASGDFCSPG EGMEILQQVC SKQLPPCNLS KEDLLQNPYF SKLLLNLSQH VDESGLSLTL AKEQAQAWKE VRLHKTTWLR SEILHRVIQ ELLVDYYVKI QDTNVTSEDK KDFVWMRARL QQEVEEQLKK KCFTLLCYYD PNSDADSETV KAAKVWKLAE V LVGEQQQC QDAKSQQKEQ MLLLEKKSAA YSQVLLRCLT LLQRLLQEHR LKTQSELDRI NAQYLEVKCG AMILKLRMEE LK ILSDTYT VEKVEVHRLI RDRLEGAIHL QEQDMENSRQ VLNSYEVLGE EFDRLVKEYT VLKQATENKR WALQEFSKVY R UniProtKB: HAUS augmin-like complex subunit 4 |
-Macromolecule #5: HAUS augmin-like complex subunit 5
Macromolecule | Name: HAUS augmin-like complex subunit 5 / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 71.783406 KDa |
Recombinant expression | Organism: Trichoplusia ni (cabbage looper) |
Sequence | String: MELAQEAREL GCWAVEEMGV PVAARAPEST LRRLCLGQGA DIWAYILQHV HSQRTVKKIR GNLLWYGHQD SPQVRRKLEL EAAVTRLRA EIQELDQSLE LMERDTEAQD TAMEQARQHT QDTQRRALLL RAQAGAMRRQ QHTLRDPMQR LQNQLRRLQD M ERKAKVDV ...String: MELAQEAREL GCWAVEEMGV PVAARAPEST LRRLCLGQGA DIWAYILQHV HSQRTVKKIR GNLLWYGHQD SPQVRRKLEL EAAVTRLRA EIQELDQSLE LMERDTEAQD TAMEQARQHT QDTQRRALLL RAQAGAMRRQ QHTLRDPMQR LQNQLRRLQD M ERKAKVDV TFGSLTSAAL GLEPVVLRDV RTACTLRAQF LQNLLLPQAK RGSLPTPHDD HFGTSYQQWL SSVETLLTNH PP GHVLAAL EHLAAEREAE IRSLCSGDGL GDTEISRPQA PDQSDSSQTL PSMVHLIQEG WRTVGVLVSQ RSTLLKERQV LTQ RLQGLV EEVERRVLGS SERQVLILGL RRCCLWTELK ALHDQSQELQ DAAGHRQLLL RELQAKQQRI LHWRQLVEET QEQV RLLIK GNSASKTRLC RSPGEVLALV QRKVVPTFEA VAPQSRELLR CLEEEVRHLP HILLGTLLRH RPGELKPLPT VLPSI HQLH PASPRGSSFI ALSHKLGLPP GKASELLLPA AASLRQDLLL LQDQRSLWCW DLLHMKTSLP PGLPTQELLQ IQASQE KQQ KENLGQALKR LEKLLKQALE RIPELQGIVG DWWEQPGQAA LSEELCQGLS LPQWRLRWVQ AQGALQKLCS UniProtKB: HAUS augmin-like complex subunit 5 |
-Macromolecule #6: HAUS augmin-like complex subunit 6
Macromolecule | Name: HAUS augmin-like complex subunit 6 / type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 50.250039 KDa |
Recombinant expression | Organism: Trichoplusia ni (cabbage looper) |
Sequence | String: MSSASVTAFE KEHLWMYLQA LGFEPGPATI ACGKIVSHTH LGVNMFDKLN RDAFHIISYF LFQVLDQSLT KEVFKFCWPP FDQKSDTEF RKHCCEWIKR ISGECGSSFP QVVGSLFLSP GGPKFIHLMY HFARFVAMKY IKSNSKNSSH HFVETFNIKP Q DLHKCIAR ...String: MSSASVTAFE KEHLWMYLQA LGFEPGPATI ACGKIVSHTH LGVNMFDKLN RDAFHIISYF LFQVLDQSLT KEVFKFCWPP FDQKSDTEF RKHCCEWIKR ISGECGSSFP QVVGSLFLSP GGPKFIHLMY HFARFVAMKY IKSNSKNSSH HFVETFNIKP Q DLHKCIAR CHFARSRFLQ ILQRQDCVTQ KYQENAQLSV KQVRNLRSEC IGLENQIKKM EPYDDHSNME EKIQKVRSLW AS VNETLMF LEKEREVVSS VLSLVNQYAL DGTNVAINIP RLLLDKIEKQ MFQLHIGNVY EAGKLNLLTV IQLLNEVLKV MKY ERCQAD QARLTVDLHY LEKETKFQKE RLSDLKHMRY RIKDDLTTIR HSVVEKQGEW HKKWKEFLGL SPFSLIKGWT PSVD LLPPM SPLSFDPASE EVYAKSILCQ YPASLPD UniProtKB: HAUS augmin-like complex subunit 6 |
-Macromolecule #7: HAUS augmin-like complex subunit 7
Macromolecule | Name: HAUS augmin-like complex subunit 7 / type: protein_or_peptide / ID: 7 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 40.819152 KDa |
Recombinant expression | Organism: Trichoplusia ni (cabbage looper) |
Sequence | String: MGGARLGARN MAGQDAGCGR GGDDYSEDEG DSSVSRAAVE VFGKLKDLNC PFLEGLYITE PKTIQELLCS PSEYRLEILE WMCTRVWPS LQDRFSSLKG VPTEVKIQEM TKLGHELMLC APDDQELLKG CACAQKQLHF MDQLLDTIRS LTIGCSSCSS L MEHFEDTR ...String: MGGARLGARN MAGQDAGCGR GGDDYSEDEG DSSVSRAAVE VFGKLKDLNC PFLEGLYITE PKTIQELLCS PSEYRLEILE WMCTRVWPS LQDRFSSLKG VPTEVKIQEM TKLGHELMLC APDDQELLKG CACAQKQLHF MDQLLDTIRS LTIGCSSCSS L MEHFEDTR EKNEALLGEL FSSPHLQMLL NPECDPWPLD MQPLLNKQSD DWQWASASAK SEEEEKLAEL ARQLQESAAK LH ALRTEYF AQHEQGAAAG AADISTLDQK LRLVTSDFHQ LILAFLQVYD DELGECCQRP GPDLHPCGPI IQATHQNLTS YSQ LLQVVM AVADTSAKAV ETVKKQQGEQ ICWGGSSSVM SLATKMNELM EK UniProtKB: HAUS augmin-like complex subunit 7 |
-Macromolecule #8: HAUS augmin-like complex subunit 8
Macromolecule | Name: HAUS augmin-like complex subunit 8 / type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 44.92243 KDa |
Recombinant expression | Organism: Trichoplusia ni (cabbage looper) |
Sequence | String: MADSSGRGAG KPATGPTNSS SAKKKDKRVQ GGRVIESRYL QYEKKTTQKA PAGDGSQTRG KMSEGGRKSS LLQKSKADSS GVGKGDLQS TLLEGHGTAP PDLDLSAIND KSIVKKTPQL AKTISKKPES TSFSAPRKKS PDLSEAMEMM ESQTLLLTLL S VKMENNLA ...String: MADSSGRGAG KPATGPTNSS SAKKKDKRVQ GGRVIESRYL QYEKKTTQKA PAGDGSQTRG KMSEGGRKSS LLQKSKADSS GVGKGDLQS TLLEGHGTAP PDLDLSAIND KSIVKKTPQL AKTISKKPES TSFSAPRKKS PDLSEAMEMM ESQTLLLTLL S VKMENNLA EFERRAEKNL LIMCKEKEKL QKKAHELKRR LLLSQRKREL ADVLDAQIEM LSPFEAVATR FKEQYRTFAT AL DTTRHEL PVRSIHLEGD GQQLLDALQH ELVTTQRLLG ELDVGDSEEN VQVLDLLSEL KDVTAKKDLE LRRSFAQVLE LSA EASKEA ALANQEVWEE TQGMAPPSRW YFNQDSACRE SGGAPKNTPL SEDDNPGASS APAQATFISP SEDFSSSSQA EVPP SLSRS GRDLS UniProtKB: HAUS augmin-like complex subunit 8 |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 0.05 mg/mL | |||||||||||||||
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Buffer | pH: 8 Component:
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Grid | Model: Quantifoil R2/2 / Material: GOLD / Support film - Material: CARBON / Support film - topology: CONTINUOUS / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 30 sec. / Pretreatment - Atmosphere: AIR | |||||||||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK III |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Specialist optics | Phase plate: VOLTA PHASE PLATE / Energy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV |
Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: SUPER-RESOLUTION / Average electron dose: 50.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 100.0 µm / Calibrated defocus max: 1.5 µm / Calibrated defocus min: 0.8 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 1.5 µm / Nominal defocus min: 0.8 µm / Nominal magnification: 64000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |