EMDB-11888: Recombinant human gTuRC

Basic information

• Entry: Database: EMDB / ID: EMD-11888
• Title: Recombinant human gTuRC
• Map data: Consensus map

Sample

• Complex: Recombinant human gamma-tubulin ring complex
  • Protein or peptide: Tubulin gamma-1 chain
  • Protein or peptide: Gamma-tubulin complex component 3
  • Protein or peptide: Gamma-tubulin complex component 6
  • Protein or peptide: Mitotic-spindle organizing protein 1
  • Protein or peptide: Actin, cytoplasmic 1
  • Protein or peptide: Gamma-tubulin complex component 2
  • Protein or peptide: Gamma-tubulin complex component 4
  • Protein or peptide: Gamma-tubulin complex component 5
• Ligand: GUANOSINE-5'-DIPHOSPHATE

• Keywords: microtubule organizing center / microtubule / gamma-tubulin ring complex / gamma-tubulin small complex / spindle organization / microtubule nucleation / CELL CYCLE

Function / homology

Function:

microtubule nucleation by interphase microtubule organizing center / gamma-tubulin complex localization / microtubule nucleator activity / positive regulation of norepinephrine uptake / polar microtubule / interphase microtubule organizing center / gamma-tubulin complex / gamma-tubulin ring complex / mitotic spindle microtubule / meiotic spindle organization / bBAF complex / cellular response to cytochalasin B / Formation of the embryonic stem cell BAF (esBAF) complex / npBAF complex / brahma complex / nBAF complex / Formation of the canonical BAF (cBAF) complex / regulation of transepithelial transport / morphogenesis of a polarized epithelium / microtubule nucleation / Formation of the polybromo-BAF (pBAF) complex / Formation of neuronal progenitor and neuronal BAF (npBAF and nBAF) / structural constituent of postsynaptic actin cytoskeleton / Formation of annular gap junctions / Formation of the dystrophin-glycoprotein complex (DGC) / GBAF complex / Gap junction degradation / Formation of the non-canonical BAF (ncBAF) complex / protein localization to adherens junction / regulation of G0 to G1 transition / Cell-extracellular matrix interactions / dense body / Folding of actin by CCT/TriC / Tat protein binding / postsynaptic actin cytoskeleton / gamma-tubulin binding / RSC-type complex / Regulation of CDH1 Function / regulation of double-strand break repair / non-motile cilium / Prefoldin mediated transfer of substrate to CCT/TriC / regulation of nucleotide-excision repair / Adherens junctions interactions / adherens junction assembly / RHOF GTPase cycle / apical protein localization / Sensory processing of sound by outer hair cells of the cochlea / tight junction / regulation of mitotic metaphase/anaphase transition / SWI/SNF complex / Sensory processing of sound by inner hair cells of the cochlea / positive regulation of T cell differentiation / Interaction between L1 and Ankyrins / apical junction complex / single fertilization / maintenance of blood-brain barrier / positive regulation of stem cell population maintenance / regulation of norepinephrine uptake / transporter regulator activity / NuA4 histone acetyltransferase complex / pericentriolar material / positive regulation of double-strand break repair / Recycling pathway of L1 / cell leading edge / Regulation of MITF-M-dependent genes involved in pigmentation / cortical cytoskeleton / ciliary transition zone / establishment or maintenance of cell polarity / microtubule organizing center / mitotic sister chromatid segregation / nitric-oxide synthase binding / brush border / EPH-ephrin mediated repulsion of cells / mitotic spindle assembly / negative regulation of cell differentiation / regulation of synaptic vesicle endocytosis / positive regulation of myoblast differentiation / RHO GTPases Activate WASPs and WAVEs / kinesin binding / regulation of protein localization to plasma membrane / RHO GTPases activate IQGAPs / positive regulation of double-strand break repair via homologous recombination / cytoplasmic microtubule / regulation of G1/S transition of mitotic cell cycle / spindle assembly / cytoplasmic microtubule organization / axonogenesis / cytoskeleton organization / EPHB-mediated forward signaling / 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 / centriole / substantia nigra development / Anchoring of the basal body to the plasma membrane / AURKA Activation by TPX2 / calyx of Held / nitric-oxide synthase regulator activity / mitotic spindle organization

Domain/homology:

Mitotic-spindle organizing protein 1 / Mitotic-spindle organizing gamma-tubulin ring associated / : / Gamma-tubulin complex component 6, N-terminal / Gamma-tubulin complex component 6 N-terminus / Gamma tubulin / Gamma tubulin complex component, C-terminal / Gamma-tubulin complex component, C-terminal domain superfamily / Gamma tubulin complex component C-terminal / Gamma-tubulin complex component protein / Gamma tubulin complex component protein, N-terminal / Gamma tubulin complex component N-terminal / Actins signature 1. / Actin, conserved site / Actins signature 2. / Actin/actin-like conserved site / Actins and actin-related proteins signature. / Tubulin / Tubulin, C-terminal / Tubulin C-terminal domain / Tubulin, conserved site / Tubulin subunits alpha, beta, and gamma signature. / Actin / Actin family / Actin / Tubulin/FtsZ family, C-terminal domain / Tubulin/FtsZ-like, C-terminal domain / Tubulin/FtsZ, C-terminal / Tubulin/FtsZ, 2-layer sandwich domain / Tubulin/FtsZ family, GTPase domain / Tubulin/FtsZ family, GTPase domain / Tubulin/FtsZ, GTPase domain / Tubulin/FtsZ, GTPase domain superfamily / ATPase, nucleotide binding domain

Component:

Tubulin gamma-1 chain / Actin, cytoplasmic 1 / Mitotic-spindle organizing protein 1 / Gamma-tubulin complex component 3 / Gamma-tubulin complex component 6 / Gamma-tubulin complex component 5 / Gamma-tubulin complex component 2 / Gamma-tubulin complex component 4

• Biological species: Homo sapiens (human)
• Method: single particle reconstruction / cryo EM / Resolution: 4.13 Å
• Authors: Serna M / Fernandez-Leiro R

Funding support

Spain, 3 items

Organization	Grant number	Country
Spanish Ministry of Science, Innovation, and Universities	SAF2017-82632-P	Spain
Other government	Y2018/BIO4747	Spain
Other government	P2018/NMT4443	Spain

• Citation: Journal: Sci Adv / Year: 2020; Title: Assembly of the asymmetric human γ-tubulin ring complex by RUVBL1-RUVBL2 AAA ATPase.; Authors: Fabian Zimmermann / Marina Serna / Artur Ezquerra / Rafael Fernandez-Leiro / Oscar Llorca / Jens Luders / ; Abstract: The microtubule nucleator γ-tubulin ring complex (γTuRC) is essential for the function of microtubule organizing centers such as the centrosome. Since its discovery over two decades ago, γTuRC has evaded in vitro reconstitution and thus detailed structure-function studies. Here, we show that a complex of RuvB-like protein 1 (RUVBL1) and RUVBL2 "RUVBL" controls assembly and composition of γTuRC in human cells. Likewise, RUVBL assembles γTuRC from a minimal set of core subunits in a heterologous coexpression system. RUVBL interacts with γTuRC subcomplexes but is not part of fully assembled γTuRC. Purified, reconstituted γTuRC has nucleation activity and resembles native γTuRC as revealed by its cryo-electron microscopy (cryo-EM) structure at ~4.0-Å resolution. We further use cryo-EM to identify features that determine the intricate, higher-order γTuRC architecture. Our work finds RUVBL as an assembly factor that regulates γTuRC in cells and allows production of recombinant γTuRC for future in-depth mechanistic studies.

History

Deposition	Oct 26, 2020	-
Header (metadata) release	Jan 20, 2021	-
Map release	Jan 20, 2021	-
Update	May 1, 2024	-
Current status	May 1, 2024	Processing site: PDBe / Status: Released

Map

• File: Download / File: emd_11888.map.gz / Format: CCP4 / Size: 226.3 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
• Annotation: Consensus map
• Voxel size: X=Y=Z: 1.40862 Å

Density

Contour Level	By AUTHOR: 0.007 / Movie #1: 0.012
Minimum - Maximum	-0.00815065 - 0.055173773
Average (Standard dev.)	-0.00011615107 (±0.002606626)

• Symmetry: Space group: 1

Details

EMDB XML:

Map geometry	Axis order X Y Z Origin 0 0 0 Dimensions 390 390 390 Spacing 390 390 390
Cell	A=B=C: 549.35986 Å α=β=γ: 90.0 °

CCP4 map header:

mode	Image stored as Reals
Å/pix. X/Y/Z	1.4086153846154	1.4086153846154	1.4086153846154
M x/y/z	390	390	390
origin x/y/z	0.000	0.000	0.000
length x/y/z	549.360	549.360	549.360
α/β/γ	90.000	90.000	90.000
MAP C/R/S	1	2	3
start NC/NR/NS	0	0	0
NC/NR/NS	390	390	390
D min/max/mean	-0.008	0.055	-0.000

Supplemental data

Additional map: Subvolume positions 1 to 3 half map 2

• File: emd_11888_additional_1.map
• Annotation: Subvolume positions 1 to 3 half map 2

Additional map: Subvolume positions 5 to 7 half map 1

• File: emd_11888_additional_10.map
• Annotation: Subvolume positions 5 to 7 half map 1

Additional map: Subvolume positions 4 to 6 half map 2

• File: emd_11888_additional_11.map
• Annotation: Subvolume positions 4 to 6 half map 2

Additional map: Subvolume positions 5 to 7

• File: emd_11888_additional_12.map
• Annotation: Subvolume positions 5 to 7

Additional map: Subvolume positions 5 to 7 half map 2

• File: emd_11888_additional_13.map
• Annotation: Subvolume positions 5 to 7 half map 2

Additional map: Subvolume positions 4 to 6 half map 1

• File: emd_11888_additional_14.map
• Annotation: Subvolume positions 4 to 6 half map 1

Additional map: Subvolume positions 4 to 6

• File: emd_11888_additional_15.map
• Annotation: Subvolume positions 4 to 6

Additional map: Subvolume positions 9 to 11 half map 1

• File: emd_11888_additional_16.map
• Annotation: Subvolume positions 9 to 11 half map 1

Additional map: Subvolume positions 9 to 11 half map 2

• File: emd_11888_additional_17.map
• Annotation: Subvolume positions 9 to 11 half map 2

Additional map: Subvolume positions 9 to 11

• File: emd_11888_additional_18.map
• Annotation: Subvolume positions 9 to 11

Additional map: Subvolume positions 8 to 10 half map 2

• File: emd_11888_additional_19.map
• Annotation: Subvolume positions 8 to 10 half map 2

Additional map: Subvolume positions 1 to 3 half map 1

• File: emd_11888_additional_2.map
• Annotation: Subvolume positions 1 to 3 half map 1

Additional map: Subvolume positions 7 to 9 half map 1

• File: emd_11888_additional_20.map
• Annotation: Subvolume positions 7 to 9 half map 1

Additional map: Subvolume positions 7 to 9 half map 2

• File: emd_11888_additional_21.map
• Annotation: Subvolume positions 7 to 9 half map 2

Additional map: Subvolume positions 8 to 10

• File: emd_11888_additional_22.map
• Annotation: Subvolume positions 8 to 10

Additional map: Subvolume positions 8 to 10 half map 1

• File: emd_11888_additional_23.map
• Annotation: Subvolume positions 8 to 10 half map 1

Additional map: Subvolume positions 6 to 8 half map 2

• File: emd_11888_additional_24.map
• Annotation: Subvolume positions 6 to 8 half map 2

Additional map: Subvolume positions 6 to 8

• File: emd_11888_additional_25.map
• Annotation: Subvolume positions 6 to 8

Additional map: Subvolume positions 6 to 8 half map 1

• File: emd_11888_additional_26.map
• Annotation: Subvolume positions 6 to 8 half map 1

Additional map: Subvolume positions 7 to 9

• File: emd_11888_additional_27.map
• Annotation: Subvolume positions 7 to 9

Additional map: Subvolume positions 11 to 14 half map 1

• File: emd_11888_additional_28.map
• Annotation: Subvolume positions 11 to 14 half map 1

Additional map: Subvolume positions 11 to 14 half map 2

• File: emd_11888_additional_29.map
• Annotation: Subvolume positions 11 to 14 half map 2

Additional map: Subvolume positions 1 to 3

• File: emd_11888_additional_3.map
• Annotation: Subvolume positions 1 to 3

Additional map: Subvolume positions 11 to 14

• File: emd_11888_additional_30.map
• Annotation: Subvolume positions 11 to 14

Additional map: Subvolume positions 10 to 12 half map 2

• File: emd_11888_additional_31.map
• Annotation: Subvolume positions 10 to 12 half map 2

Additional map: Subvolume positions 10 to 12 half map 1

• File: emd_11888_additional_32.map
• Annotation: Subvolume positions 10 to 12 half map 1

Additional map: Subvolume positions 10 to 12

• File: emd_11888_additional_33.map
• Annotation: Subvolume positions 10 to 12

Additional map: Subvolume luminal bridge half map 2

• File: emd_11888_additional_34.map
• Annotation: Subvolume luminal bridge half map 2

Additional map: Subvolume luminal bridge

• File: emd_11888_additional_35.map
• Annotation: Subvolume luminal bridge

Additional map: Subvolume luminal bridge half map 1

• File: emd_11888_additional_36.map
• Annotation: Subvolume luminal bridge half map 1

Additional map: Subvolume positions 2 to 4

• File: emd_11888_additional_4.map
• Annotation: Subvolume positions 2 to 4

Additional map: Subvolume positions 2 to 4 half map 2

• File: emd_11888_additional_5.map
• Annotation: Subvolume positions 2 to 4 half map 2

Additional map: Subvolume positions 2 to 4 half map 1

• File: emd_11888_additional_6.map
• Annotation: Subvolume positions 2 to 4 half map 1

Additional map: Subvolume positions 3 to 5 half map 1

• File: emd_11888_additional_7.map
• Annotation: Subvolume positions 3 to 5 half map 1

Additional map: Subvolume positions 3 to 5

• File: emd_11888_additional_8.map
• Annotation: Subvolume positions 3 to 5

Additional map: Subvolume positions 3 to 5 half map 2

• File: emd_11888_additional_9.map
• Annotation: Subvolume positions 3 to 5 half map 2

Half map: Consensus half map 2

• File: emd_11888_half_map_1.map
• Annotation: Consensus half map 2

Half map: Consensus half map 1

• File: emd_11888_half_map_2.map
• Annotation: Consensus half map 1

Sample components

Entire : Recombinant human gamma-tubulin ring complex

• Entire: Name: Recombinant human gamma-tubulin ring complex

Components

• Complex: Recombinant human gamma-tubulin ring complex
  • Protein or peptide: Tubulin gamma-1 chain
  • Protein or peptide: Gamma-tubulin complex component 3
  • Protein or peptide: Gamma-tubulin complex component 6
  • Protein or peptide: Mitotic-spindle organizing protein 1
  • Protein or peptide: Actin, cytoplasmic 1
  • Protein or peptide: Gamma-tubulin complex component 2
  • Protein or peptide: Gamma-tubulin complex component 4
  • Protein or peptide: Gamma-tubulin complex component 5
• Ligand: GUANOSINE-5'-DIPHOSPHATE

Supramolecule #1: Recombinant human gamma-tubulin ring complex

• Supramolecule: Name: Recombinant human gamma-tubulin ring complex / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#8
• Source (natural): Organism: Homo sapiens (human)

Macromolecule #1: Tubulin gamma-1 chain

• Macromolecule: Name: Tubulin gamma-1 chain / type: protein_or_peptide / ID: 1 / Number of copies: 14 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 50.741297 KDa
• Recombinant expression: Organism: Spodoptera frugiperda (fall armyworm)

Sequence

String:

MPREIITLQL GQCGNQIGFE FWKQLCAEHG ISPEGIVEEF ATEGTDRKDV FFYQADDEHY IPRAVLLDLE PRVIHSILNS PYAKLYNPE NIYLSEHGGG AGNNWASGFS QGEKIHEDIF DIIDREADGS DSLEGFVLCH SIAGGTGSGL GSYLLERLND R YPKKLVQT YSVFPNQDEM SDVVVQPYNS LLTLKRLTQN ADCVVVLDNT ALNRIATDRL HIQNPSFSQI NQLVSTIMSA ST TTLRYPG YMNNDLIGLI ASLIPTPRLH FLMTGYTPLT TDQSVASVRK TTVLDVMRRL LQPKNVMVST GRDRQTNHCY IAI LNIIQG EVDPTQVHKS LQRIRERKLA NFIPWGPASI QVALSRKSPY LPSAHRVSGL MMANHTSISS LFERTCRQYD KLRK REAFL EQFRKEDMFK DNFDEMDTSR EIVQQLIDEY HAATRPDYIS WG

UniProtKB: Tubulin gamma-1 chain

Macromolecule #2: Gamma-tubulin complex component 3

• Macromolecule: Name: Gamma-tubulin complex component 3 / type: protein_or_peptide / ID: 2 / Number of copies: 6 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 103.710102 KDa
• Recombinant expression: Organism: Spodoptera frugiperda (fall armyworm)

Sequence

String:

MATPDQKSPN VLLQNLCCRI LGRSEADVAQ QFQYAVRVIG SNFAPTVERD EFLVAEKIKK ELIRQRREAD AALFSELHRK LHSQGVLKN KWSILYLLLS LSEDPRRQPS KVSSYATLFA QALPRDAHST PYYYARPQTL PLSYQDRSAQ SAQSSGSVGS S GISSIGLC ALSGPAPAPQ SLLPGQSNQA PGVGDCLRQQ LGSRLAWTLT ANQPSSQATT SKGVPSAVSR NMTRSRREGD TG GTMEITE AALVRDILYV FQGIDGKNIK MNNTENCYKV EGKANLSRSL RDTAVRLSEL GWLHNKIRRY TDQRSLDRSF GLV GQSFCA ALHQELREYY RLLSVLHSQL QLEDDQGVNL GLESSLTLRR LLVWTYDPKI RLKTLAALVD HCQGRKGGEL ASAV HAYTK TGDPYMRSLV QHILSLVSHP VLSFLYRWIY DGELEDTYHE FFVASDPTVK TDRLWHDKYT LRKSMIPSFM TMDQS RKVL LIGKSINFLH QVCHDQTPTT KMIAVTKSAE SPQDAADLFT DLENAFQGKI DAAYFETSKY LLDVLNKKYS LLDHMQ AMR RYLLLGQGDF IRHLMDLLKP ELVRPATTLY QHNLTGILET AVRATNAQFD SPEILRRLDV RLLEVSPGDT GWDVFSL DY HVDGPIATVF TRECMSHYLR VFNFLWRAKR MEYILTDIRK GHMCNAKLLR NMPEFSGVLH QCHILASEMV HFIHQMQY Y ITFEVLECSW DELWNKVQQA QDLDHIIAAH EVFLDTIISR CLLDSDSRAL LNQLRAVFDQ IIELQNAQDA IYRAALEEL QRRLQFEEKK KQREIEGQWG VTAAEEEEEN KRIGEFKESI PKMCSQLRIL THFYQGIVQQ FLVLLTTSSD ESLRFLSFRL DFNEHYKAR EPRLRVSLGT RGRRSSHT

UniProtKB: Gamma-tubulin complex component 3

Macromolecule #3: Gamma-tubulin complex component 6

• Macromolecule: Name: Gamma-tubulin complex component 6 / type: protein_or_peptide / ID: 3 / Number of copies: 2 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 200.733641 KDa
• Recombinant expression: Organism: Spodoptera frugiperda (fall armyworm)

Sequence

String:

MASITQLFDD LCEALLPAAK THLGQRSVNR KRAKRSLKKV AYNALFTNLF QDETQQLQPD MSKLPARNKI LMLSFDLRVG GLGPKADRL EELVEELEAA PCCPLLEVGS VLDLLVQLAG SGPPQVLPRK RDYFLNNKHV GRNVPYSGYD CDDLSVFEMD V QSLISREE CLCHSMIQET LQVMEAAPGT GLPTVGLFSF GDPCGDRFER DTRVSLFGAL VHSRTYDMDV RLGLPPVPDN AD LSGLAIK VPPSVDQWED EGFQSASNLT PDSQSEPSVT PDVDLWEAAL TYEASKRRCW ERVGCPPGHR EEPYLTEAGR DAF DKFCRL HQGELQLLAG GVLQAPQPVL VKECELVKDV LNVLIGVVSA TFSLCQPAQA FVVKRGVHVS GASPESISSL LSEV AEYGT CYTRLSHFSL QPVLDSLYSK GLVFQAFTSG LRRYLQYYRA CVLSTPPTLS LLTIGFLFKK LGRQLRYLAE LCGVG AVLP GTCGGGPRAA FPTGVKLLSY LYQEALHNCS NEHYPVLLSL LKTSCEPYTR FIHDWVYSGV FRDAYGEFMI QVNHEY LSF RDKLYWTHGY VLISKEVEDC VPVFLKHIAH DIYVCGKTIN LLKLCCPRHY LCWSDVPVPR ISVIFSLEEL KEIEKDC AV YVGRMERVAR HSSVSKEEKE LRMEIAKQEL IAHAREAASR VLSALSDRQM SERMALDARK REQFQRLKEQ FVKDQERR Q AARQEELDDD FSYARELRDR ERRLKSLEEE LERKARQALV DHYSKLSAEA ARREQKALWR IQRHRLESAR LRFLLEDEK HIQEMLKAVS EAHQPQEPPD VLLSVHPQVT SPGPEHPEGG QGCDSGSAEQ HSPAWDGWNR PGLLTPQPLK PLAVGAGGRG LQQAEGARP FSDSLSIGDF LPVGPGAEPS VQTGMVPLLE VALQTINLDL PPSAPGEAPA AASTQPSRPQ EYDFSTVLRP A VATSPAPG PLQAAECSLG SSGLQLWEDS CGKMDACGSA SRETLLPSHP PRRAALEEGS SQPTERLFGQ VSGGGLPTGD YA SEIAPTR PRWNTHGHVS DASIRVGENV SDVAPTQPRW NTHGHVSNAS ISLGESVSDV APTRPRWNIH GHVSNASIRV GEN VSDVAP TRPRWNTHGH VSNASIRVGE NVSDVAPTRP RWNTHGHVSD ASISLGESVS DMAPARPRWN THGHVSDASI SLGE SVSDM APTRPRWNTH GHVSDTSIRV GENVSDVAPI RSRCNTHGHV SDASISLGEP VSDVVSTRPR WNTHVPIPPP HMVLG ALSP EAEPNTPRPQ QSPPGHTSQS ALSLGAQSTV LDCGPRLPVE VGPSLSSPSS GCGEGSISVG ENVSDVAPTQ PWWPNT PGD SVSEELGPGR SGDTEDLSPN WPLNSQEDTA AQSSPGRGEE AEASAAEAQG GEQAYLAGLA GQYHLERYPD SYESMSE PP IAHLLRPVLP RAFAFPVDPQ VQSAADETAV QLSELLTLPV LMKRSITAPL AAHISLVNKA AVDYFFVELH LEAHYEAL R HFLLMEDGEF AQSLSDLLFE KLGAGQTPGE LLNPLVLNSV LSKALQCSLH GDTPHASNLS LALKYLPEVF APNAPDVLS CLELRYKVDW PLNIVITEGC VSKYSGVFSF LLQLKLMMWA LKDVCFHLKR TALLSHMAGS VQFRQLQLFK HEMQHFVKVI QGYIANQIL HVTWCEFRAR LATVGDLEEI QRAHAEYLHK AVFRGLLTEK AAPVMNVIHS IFSLVLKFRS QLISQAWGPP G GPRGAEHP NFALMQQSYN TFKYYSHFLF KVVTKLVNRG YQPHLEDFLL RINFNNYYQD A

UniProtKB: Gamma-tubulin complex component 6

Macromolecule #4: Mitotic-spindle organizing protein 1

• Macromolecule: Name: Mitotic-spindle organizing protein 1 / type: protein_or_peptide / ID: 4 / Number of copies: 2 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 8.485724 KDa
• Recombinant expression: Organism: Spodoptera frugiperda (fall armyworm)

Sequence

String:

MASSSGAGAA AAAAAANLNA VRETMDVLLE ISRILNTGLD METLSICVRL CEQGINPEAL SSVIKELRKA TEALKAAENM TS

UniProtKB: Mitotic-spindle organizing protein 1

Macromolecule #5: Actin, cytoplasmic 1

• Macromolecule: Name: Actin, cytoplasmic 1 / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 41.723527 KDa
• Recombinant expression: Organism: Spodoptera frugiperda (fall armyworm)

Sequence

String:

DEEVAALVVD NGSGMCKAGF AGDDAPRAVF PSIVGRPRHQ GVMVGMGQKD SYVGDEAQSK RGILTLKYPI EHGIVTNWDD MEKIWHHTF YNELRVAPEE HPVLLTEAPL NPKANREKMT QIMFETFNTP AMYVAIQAVL SLYASGRTTG IVLDSGDGVS H TVPIYEGY ALPHAILRLD LAGRDLTDYL MKILTERGYS FTTTEEREIV RDIKEKLCYV ALDFEQEMAT AASSSSLEKS YE LKDGQVI TIGNERFRCP EALFQPSFLG MEACGIHETT YNSIMKCDVD IRKDLYANTV LSGGTTMYPG IADRMQKEIT ALA PSTMKI KIIAPPERKY SVWIGGSILA SLSTFQQMWI SKQEYDESGP SIVHRKCF

UniProtKB: Actin, cytoplasmic 1

Macromolecule #6: Gamma-tubulin complex component 2

• Macromolecule: Name: Gamma-tubulin complex component 2 / type: protein_or_peptide / ID: 6 / Number of copies: 5 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 102.666953 KDa
• Recombinant expression: Organism: Spodoptera frugiperda (fall armyworm)

Sequence

String:

MSEFRIHHDV NELLSLLRVH GGDGAEVYID LLQKNRTPYV TTTVSAHSAK VKIAEFSRTP EDFLKKYDEL KSKNTRNLDP LVYLLSKLT EDKETLQYLQ QNAKERAELA AAAVGSSTTS INVPAAASKI SMQELEELRK QLGSVATGST LQQSLELKRK M LRDKQNKK NSGQHLPIFP AWVYERPALI GDFLIGAGIS TDTALPIGTL PLASQESAVV EDLLYVLVGV DGRYVSAQPL AG RQSRTFL VDPNLDLSIR ELVHRILPVA ASYSAVTRFI EEKSSFEYGQ VNHALAAAMR TLVKEHLILV SQLEQLHRQG LLS LQKLWF YIQPAMRTMD ILASLATSVD KGECLGGSTL SLLHDRSFSY TGDSQAQELC LYLTKAASAP YFEVLEKWIY RGII HDPYS EFMVEEHELR KERIQEDYND KYWDQRYTIV QQQIPSFLQK MADKILSTGK YLNVVRECGH DVTCPVAKEI IYTLK ERAY VEQIEKAFNY ASKVLLDFLM EEKELVAHLR SIKRYFLMDQ GDFFVHFMDL AEEELRKPVE DITPPRLEAL LELALR MST ANTDPFKDDL KIDLMPHDLI TQLLRVLAIE TKQEKAMAHA DPTELALSGL EAFSFDYIVK WPLSLIINRK ALTRYQM LF RHMFYCKHVE RQLCSVWISN KTAKQHSLHS AQWFAGAFTL RQRMLNFVQN IQYYMMFEVM EPTWHILEKN LKSASNID D VLGHHTGFLD TCLKDCMLTN PELLKVFSKL MSVCVMFTNC MQKFTQSMKL DGELGGQTLE HSTVLGLPAG AEERARKEL ARKHLAEHAD TVQLVSGFEA TINKFDKNFS AHLLDLLARL SIYSTSDCEH GMASVISRLD FNGFYTERLE RLSAERSQKA TPQVPVLRG PPAPAPRVAV TAQ

UniProtKB: Gamma-tubulin complex component 2

Macromolecule #7: Gamma-tubulin complex component 4

• Macromolecule: Name: Gamma-tubulin complex component 4 / type: protein_or_peptide / ID: 7 / Number of copies: 2 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 76.179969 KDa
• Recombinant expression: Organism: Spodoptera frugiperda (fall armyworm)

Sequence

String:

MIHELLLALS GYPGSIFTWN KRSGLQVSQD FPFLHPSETS VLNRLCRLGT DYIRFTEFIE QYTGHVQQQD HHPSQQGQGG LHGIYLRAF CTGLDSVLQP YRQALLDLEQ EFLGDPHLSI SHVNYFLDQF QLLFPSVMVV VEQIKSQKIH GCQILETVYK H SCGGLPPV RSALEKILAV CHGVMYKQLS AWMLHGLLLD QHEEFFIKQG PSSGNVSAQP EEDEEDLGIG GLTGKQLREL QD LRLIEEE NMLAPSLKQF SLRVEILPSY IPVRVAEKIL FVGESVQMFE NQNVNLTRKG SILKNQEDTF AAELHRLKQQ PLF SLVDFE QVVDRIRSTV AEHLWKLMVE ESDLLGQLKI IKDFYLLGRG ELFQAFIDTA QHMLKTPPTA VTEHDVNVAF QQSA HKVLL DDDNLLPLLH LTIEYHGKEH KADATQAREG PSRETSPREA PASGWAALGL SYKVQWPLHI LFTPAVLEKY NVVFK YLLS VRRVQAELQH CWALQMQRKH LKSNQTDAIK WRLRNHMAFL VDNLQYYLQV DVLESQFSQL LHQINSTRDF ESIRLA HDH FLSNLLAQSF ILLKPVFHCL NEILDLCHSF CSLVSQNLGP LDERGAAQLS ILVKGFSRQS SLLFKILSSV RNHQINS DL AQLLLRLDYN KYYTQAGGTL GSFGM

UniProtKB: Gamma-tubulin complex component 4

Macromolecule #8: Gamma-tubulin complex component 5

• Macromolecule: Name: Gamma-tubulin complex component 5 / type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 118.467547 KDa
• Recombinant expression: Organism: Spodoptera frugiperda (fall armyworm)

Sequence

String:

MARHGPPWSR LDAQQERDVR ELVRGVAGLQ DEADPNFQLA LNFAWSNFRF HRFLDVNSHK IEKTIEGIYE KFVIHSDLSK AASWKRLTE EFLNAPLPSI KEIKTDAHYS ILSLLLCLSD SPSNSSYVET PRNKEVEKKD DFDWGKYLME DEEMDIGPYM D TPNWSEES EEENDQQPLS REDSGIQVDR TPLEEQDQNR KLDPCISWKD EPDDRSWLEH HVVHQYWTAR PSQFPHSLHL HS NLAAVWD QHLYSSDPLY VPDDRVLVTE TQVIRETLWL LSGVKKLFIF QLIDGKVTVR NNIIVTHLTH SCLRSVLEQI AAY GQVVFR LQEFIDEVMG HSSESMLPGS GSVPKKSTEA PFRTYQAFMW ALYKYFISFK EELAEIEKCI INNDTTITLA IVVD KLAPR LSQLKVLHKV FSTGVAEVPP DTRNVVRASH LLNTLYKAIL EYDNVGEASE QTVSLLFSLW VETVRPYLQT VDEWI VHGH LWDGAREFII QRNKNVPVNH RDFWYATYTL YSVSEKTENE EKMSDNASAS SGSDQGPSSR QHTMVSFLKP VLKQII MAG KSMQLLKNLQ CAESTTCQAG ARDAERKSLY TLFLESVQSR LRHGEDSTPQ VLTEQQATKE NLMKMQSIAE SHLELDD VH DPLLAINFAR MYLEQSDFHE KFAGGDVCVD RSSESVTCQT FELTLRSCLY PHIDKQYLDC CGNLMQTLKK DYRLVEYL Q AMRNFFLMEG GDTMYDFYTS IFDKIREKET WQNVSFLNVQ LQEAVGQRYP EDSSRLSISF ENVDTAKKKL PVHILDGLT LSYKVPWPVD IVISLECQKI YNQVFLLLLQ IKWAKYSLDV LLFGELVSTA EKPRLKEGLI HEQDTVAQFG PQKEPVRQQI HRMFLLRVK LMHFVNSLHN YIMTRILHST GLEFQHQVEE AKDLDQLIKI HYRYLSTIHD RCLLREKVSF VKEAIMKVLN L ALMFADGW QAGLGTWRME SIEKMESDFK NCHMFLVTIL NKAVCRGSFP HLESLALSLM AGMEQS

UniProtKB: Gamma-tubulin complex component 5

Macromolecule #9: GUANOSINE-5'-DIPHOSPHATE

• Macromolecule: Name: GUANOSINE-5'-DIPHOSPHATE / type: ligand / ID: 9 / Number of copies: 14 / Formula: GDP
• Molecular weight: Theoretical: 443.201 Da

Chemical component information

ChemComp-GDP:
GUANOSINE-5'-DIPHOSPHATE / GDP, energy-carrying molecule*YM

Experimental details

Structure determination

• Method: cryo EM
• Processing: single particle reconstruction
• Aggregation state: particle

Sample preparation

• Buffer: pH: 7.5
• Vitrification: Cryogen name: ETHANE

Electron microscopy

• Microscope: FEI TITAN KRIOS
• Image recording: Film or detector model: GATAN K3 (6k x 4k) / Average electron dose: 58.0 e/Ų
• Electron beam: Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
• Electron optics: Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD
• Experimental equipment: Model: Titan Krios / Image courtesy: FEI Company

Image processing

• Startup model: Type of model: OTHER / Details: Ab initio
• Final reconstruction: Resolution.type: BY AUTHOR / Resolution: 4.13 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 105181
• Initial angle assignment: Type: MAXIMUM LIKELIHOOD
• Final angle assignment: Type: MAXIMUM LIKELIHOOD