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- EMDB-14336: Structure of the human CCAN bound to alpha satellite DNA -

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Basic information

Entry
Database: EMDB / ID: EMD-14336
TitleStructure of the human CCAN bound to alpha satellite DNA
Map dataUnsharpened map from Relion Refine3D
Sample
  • Complex: Human CCAN complex with bound alpha-satellite DNA
    • Protein or peptide: x 15 types
    • DNA: x 1 types
  • DNA: x 1 types
  • Ligand: x 1 types
Function / homology
Function and homology information


positive regulation of protein localization to kinetochore / Mis6-Sim4 complex / FANCM-MHF complex / centromere complex assembly / kinetochore organization / metaphase chromosome alignment / Fanconi anaemia nuclear complex / inner kinetochore / kinetochore binding / sex differentiation ...positive regulation of protein localization to kinetochore / Mis6-Sim4 complex / FANCM-MHF complex / centromere complex assembly / kinetochore organization / metaphase chromosome alignment / Fanconi anaemia nuclear complex / inner kinetochore / kinetochore binding / sex differentiation / CENP-A containing chromatin assembly / resolution of meiotic recombination intermediates / chordate embryonic development / negative regulation of epithelial cell apoptotic process / kinetochore assembly / replication fork processing / mitotic sister chromatid segregation / chromosome, centromeric region / centriolar satellite / chromosome organization / Amplification of signal from unattached kinetochores via a MAD2 inhibitory signal / interstrand cross-link repair / Mitotic Prometaphase / EML4 and NUDC in mitotic spindle formation / Deposition of new CENPA-containing nucleosomes at the centromere / Resolution of Sister Chromatid Cohesion / NRIF signals cell death from the nucleus / mitotic spindle organization / positive regulation of protein ubiquitination / chromosome segregation / positive regulation of epithelial cell proliferation / RHO GTPases Activate Formins / Fanconi Anemia Pathway / PKR-mediated signaling / kinetochore / nuclear matrix / Separation of Sister Chromatids / actin cytoskeleton / mitotic cell cycle / chromosome / nuclear body / cell adhesion / protein heterodimerization activity / cell division / DNA repair / apoptotic process / DNA damage response / chromatin binding / chromatin / nucleolus / regulation of DNA-templated transcription / signal transduction / DNA binding / nucleoplasm / membrane / nucleus / cytosol / cytoplasm
Similarity search - Function
Centromere protein W / CENP-W protein / Centromere protein T / Centromere kinetochore component CENP-T, N-terminal domain / Centromere kinetochore component CENP-T N-terminus / Centromere subunit L / Kinetochore complex Sim4 subunit Fta1 / Centromere protein R / Centromere protein Q / Centromere protein U ...Centromere protein W / CENP-W protein / Centromere protein T / Centromere kinetochore component CENP-T, N-terminal domain / Centromere kinetochore component CENP-T N-terminus / Centromere subunit L / Kinetochore complex Sim4 subunit Fta1 / Centromere protein R / Centromere protein Q / Centromere protein U / Centromere protein P / Centromere protein H / Kinetochore component, CENP-R / CENP-Q, a CENPA-CAD centromere complex subunit / CENP-A-nucleosome distal (CAD) centromere subunit, CENP-P / CENP-A nucleosome associated complex (NAC) subunit / Centromere protein H, C-terminal / Centromere protein Cenp-M / Centromere protein Cenp-K / Centromere protein H (CENP-H) / Centromere protein M (CENP-M) / Centromere-associated protein K / Centromere protein I / Mis6 / Centromere protein X / CENP-S/Mhf1 / CENP-S associating Centromere protein X / CENP-S protein / Centromere protein O / Cenp-O kinetochore centromere component / Centromere protein Chl4/mis15/CENP-N / Kinetochore protein CHL4 like / CENP-T/Histone H4, histone fold / Centromere kinetochore component CENP-T histone fold / Histone-fold / P-loop containing nucleoside triphosphate hydrolase
Similarity search - Domain/homology
Centromere protein X / Centromere protein R / Centromere protein W / Centromere protein P / Centromere protein U / Centromere protein Q / Centromere protein L / Centromere protein S / Centromere protein I / Centromere protein T ...Centromere protein X / Centromere protein R / Centromere protein W / Centromere protein P / Centromere protein U / Centromere protein Q / Centromere protein L / Centromere protein S / Centromere protein I / Centromere protein T / Centromere protein N / Centromere protein K / Centromere protein O / Centromere protein H / Centromere protein M
Similarity search - Component
Biological speciesHomo sapiens (human)
Methodsingle particle reconstruction / cryo EM / Resolution: 2.83 Å
AuthorsYatskevich S / Muir KW / Bellini D / Zhang Z / Yang J / Tischer T / Predin M / Dendooven T / McLaughlin SH / Barford D
Funding support United Kingdom, Germany, 3 items
OrganizationGrant numberCountry
Cancer Research UK United Kingdom
Boehringer Ingelheim Fonds (BIF) Germany
Medical Research Council (MRC, United Kingdom) United Kingdom
CitationJournal: Science / Year: 2022
Title: Structure of the human inner kinetochore bound to a centromeric CENP-A nucleosome.
Authors: Stanislau Yatskevich / Kyle W Muir / Dom Bellini / Ziguo Zhang / Jing Yang / Thomas Tischer / Masa Predin / Tom Dendooven / Stephen H McLaughlin / David Barford /
Abstract: Kinetochores assemble onto specialized centromeric CENP-A (centromere protein A) nucleosomes (CENP-A) to mediate attachments between chromosomes and the mitotic spindle. We describe cryo-electron ...Kinetochores assemble onto specialized centromeric CENP-A (centromere protein A) nucleosomes (CENP-A) to mediate attachments between chromosomes and the mitotic spindle. We describe cryo-electron microscopy structures of the human inner kinetochore constitutive centromere associated network (CCAN) complex bound to CENP-A reconstituted onto α-satellite DNA. CCAN forms edge-on contacts with CENP-A, and a linker DNA segment of the α-satellite repeat emerges from the fully wrapped end of the nucleosome to thread through the central CENP-LN channel that tightly grips the DNA. The CENP-TWSX histone-fold module further augments DNA binding and partially wraps the linker DNA in a manner reminiscent of canonical nucleosomes. Our study suggests that the topological entrapment of the linker DNA by CCAN provides a robust mechanism by which kinetochores withstand both pushing and pulling forces exerted by the mitotic spindle.
History
DepositionFeb 11, 2022-
Header (metadata) releaseApr 27, 2022-
Map releaseApr 27, 2022-
UpdateJun 1, 2022-
Current statusJun 1, 2022Processing site: PDBe / Status: Released

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Structure visualization

Supplemental images

Downloads & links

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Map

FileDownload / File: emd_14336.map.gz / Format: CCP4 / Size: 307.5 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationUnsharpened map from Relion Refine3D
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
0.85 Å/pix.
x 432 pix.
= 368.496 Å
0.85 Å/pix.
x 432 pix.
= 368.496 Å
0.85 Å/pix.
x 432 pix.
= 368.496 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 0.853 Å
Density
Contour LevelBy AUTHOR: 0.007
Minimum - Maximum-0.011673657 - 0.039002478
Average (Standard dev.)0.00016291883 (±0.0010269354)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions432432432
Spacing432432432
CellA=B=C: 368.496 Å
α=β=γ: 90.0 °

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Supplemental data

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Additional map: Sharpened map from Relion PostProcess

Fileemd_14336_additional_1.map
AnnotationSharpened map from Relion PostProcess
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

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Sample components

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Entire : Human CCAN complex with bound alpha-satellite DNA

EntireName: Human CCAN complex with bound alpha-satellite DNA
Components
  • Complex: Human CCAN complex with bound alpha-satellite DNA
    • Protein or peptide: Centromere protein H
    • Protein or peptide: Centromere protein I
    • DNA: DNA (66-MER)
    • Protein or peptide: Centromere protein K
    • Protein or peptide: Centromere protein L
    • Protein or peptide: Centromere protein MCENPM
    • Protein or peptide: Centromere protein N
    • Protein or peptide: Centromere protein OCENPO
    • Protein or peptide: Centromere protein P
    • Protein or peptide: Centromere protein Q
    • Protein or peptide: Centromere protein U
    • Protein or peptide: Centromere protein R
    • Protein or peptide: Centromere protein T
    • Protein or peptide: Centromere protein W
    • Protein or peptide: Centromere protein S
    • Protein or peptide: Centromere protein X
  • DNA: DNA (66-MER)
  • Ligand: water

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Supramolecule #1: Human CCAN complex with bound alpha-satellite DNA

SupramoleculeName: Human CCAN complex with bound alpha-satellite DNA / type: complex / Chimera: Yes / ID: 1 / Parent: 0 / Macromolecule list: #1-#16

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Macromolecule #1: Centromere protein H

MacromoleculeName: Centromere protein H / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 28.520941 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MEEQPQMQDA DEPADSGGEG RAGGPPQVAG AQAACSEDRM TLLLRLRAQT KQQLLEYKSM VDASEEKTPE QIMQEKQIEA KIEDLENEI EEVKVAFEIK KLALDRMRLS TALKKNLEKI SRQSSVLMDN MKHLLELNKL IMKSQQESWD LEEKLLDIRK K RLQLKQAS ...String:
MEEQPQMQDA DEPADSGGEG RAGGPPQVAG AQAACSEDRM TLLLRLRAQT KQQLLEYKSM VDASEEKTPE QIMQEKQIEA KIEDLENEI EEVKVAFEIK KLALDRMRLS TALKKNLEKI SRQSSVLMDN MKHLLELNKL IMKSQQESWD LEEKLLDIRK K RLQLKQAS ESKLLEIQTE KNKQKIDLDS MENSERIKII RQNLQMEIKI TTVIQHVFQN LILGSKVNWA EDPALKEIVL QL EKNVDMM

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Macromolecule #2: Centromere protein I

MacromoleculeName: Centromere protein I / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 86.820188 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MSPQKRVKNV QAQNRTSQGS SSFQTTLSAW KVKQDPSNSK NISKHGQNNP VGDYEHADDQ AEEDALQMAV GYFEKGPIKA SQNKDKTLE KHLKTVENVA WKNGLASEEI DILLNIALSG KFGNAVNTRI LKCMIPATVI SEDSVVKAVS WLCVGKCSGS T KVLFYRWL ...String:
MSPQKRVKNV QAQNRTSQGS SSFQTTLSAW KVKQDPSNSK NISKHGQNNP VGDYEHADDQ AEEDALQMAV GYFEKGPIKA SQNKDKTLE KHLKTVENVA WKNGLASEEI DILLNIALSG KFGNAVNTRI LKCMIPATVI SEDSVVKAVS WLCVGKCSGS T KVLFYRWL VAMFDFIDRK EQINLLYGFF FASLQDDALC PYVCHLLYLL TKKENVKPFR VRKLLDLQAK MGMQPHLQAL LS LYKFFAP ALISVSLPVR KKIYFKNSEN LWKTALLAVK QRNRGPSPEP LKLMLGPANV RPLKRKWNSL SVIPVLNSSS YTK ECGKKE MSLSDCLNRS GSFPLEQLQS FPQLLQNIHC LELPSQMGSV LNNSLLLHYI NCVRDEPVLL RFYYWLSQTL QEEC IWYKV NNYEHGKEFT NFLDTIIRAE CFLQEGFYSC EAFLYKSLPL WDGLCCRSQF LQLVSWIPFS SFSEVKPLLF DHLAQ LFFT STIYFKCSVL QSLKELLQNW LLWLSMDIHM KPVTNSPLET TLGGSMNSVS KLIHYVGWLS TTAMRLESNN TFLLHF ILD FYEKVCDIYI NYNLPLVVLF PPGIFYSALL SLDTSILNQL CFIMHRYRKN LTAAKKNELV QKTKSEFNFS SKTYQEF NH YLTSMVGCLW TSKPFGKGIY IDPEILEKTG VAEYKNSLNV VHHPSFLSYA VSFLLQESPE ERTVNVSSIR GKKWSWYL D YLFSQGLQGL KLFIRSSVHH SSIPRAEGIN CNNQY

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Macromolecule #4: Centromere protein K

MacromoleculeName: Centromere protein K / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 31.69607 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MNQEDLDPDS TTDVGDVTNT EEELIRECEE MWKDMEECQN KLSLIGTETL TDSNAQLSLL IMQVKCLTAE LSQWQKKTPE TIPLTEDVL ITLGKEEFQK LRQDLEMVLS TKESKNEKLK EDLEREQRWL DEQQQIMESL NVLHSELKNK VETFSESRIF N ELKTKMLN ...String:
MNQEDLDPDS TTDVGDVTNT EEELIRECEE MWKDMEECQN KLSLIGTETL TDSNAQLSLL IMQVKCLTAE LSQWQKKTPE TIPLTEDVL ITLGKEEFQK LRQDLEMVLS TKESKNEKLK EDLEREQRWL DEQQQIMESL NVLHSELKNK VETFSESRIF N ELKTKMLN IKEYKEKLLS TLGEFLEDHF PLPDRSVKKK KKNIQESSVN LITLHEMLEI LINRLFDVPH DPYVKISDSF WP PYVELLL RNGIALRHPE DPTRIRLEAF HQ

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Macromolecule #5: Centromere protein L

MacromoleculeName: Centromere protein L / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 39.039641 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MDSYSAPEST PSASSRPEDY FIGATPLQKR LESVRKQSSF ILTPPRRKIP QCSQLQEDVD PQKVAFLLHK QWTLYSLTPL YKFSYSNLK EYSRLLNAFI VAEKQKGLAV EVGEDFNIKV IFSTLLGMKG TQRDPEAFLV QIVSKSQLPS ENREGKVLWT G WFCCVFGD ...String:
MDSYSAPEST PSASSRPEDY FIGATPLQKR LESVRKQSSF ILTPPRRKIP QCSQLQEDVD PQKVAFLLHK QWTLYSLTPL YKFSYSNLK EYSRLLNAFI VAEKQKGLAV EVGEDFNIKV IFSTLLGMKG TQRDPEAFLV QIVSKSQLPS ENREGKVLWT G WFCCVFGD SLLETVSEDF TCLPLFLANG AESNTAIIGT WFQKTFDCYF SPLAINAFNL SWMAAMWTAC KMDHYVATTE FL WSVPCSP QSLDISFAIH PEDAKALWDS VHKTPGEVTQ EEVDLFMDCL YSHFHRHFKI HLSATRLVRV STSVASAHTD GKI KILCHK YLIGVLAYLT ELAIFQIE

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Macromolecule #6: Centromere protein M

MacromoleculeName: Centromere protein M / type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 19.761945 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString:
MSVLRPLDKL PGLNTATILL VGTEDALLQQ LADSMLKEDC ASELKVHLAK SLPLPSSVNR PRIDLIVFVV NLHSKYSLQN TEESLRHVD ASFFLGKVCF LATGAGRESH CSIHRHTVVK LAHTYQSPLL YCDLEVEGFR ATMAQRLVRV LQICAGHVPG V SALNLLSL LRSSEGPSLE DL

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Macromolecule #7: Centromere protein N

MacromoleculeName: Centromere protein N / type: protein_or_peptide / ID: 7 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 39.609551 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MDETVAEFIK RTILKIPMNE LTTILKAWDF LSENQLQTVN FRQRKESVVQ HLIHLCEEKR ASISDAALLD IIYMQFHQHQ KVWEVFQMS KGPGEDVDLF DMKQFKNSFK KILQRALKNV TVSFRETEEN AVWIRIAWGT QYTKPNQYKP TYVVYYSQTP Y AFTSSSML ...String:
MDETVAEFIK RTILKIPMNE LTTILKAWDF LSENQLQTVN FRQRKESVVQ HLIHLCEEKR ASISDAALLD IIYMQFHQHQ KVWEVFQMS KGPGEDVDLF DMKQFKNSFK KILQRALKNV TVSFRETEEN AVWIRIAWGT QYTKPNQYKP TYVVYYSQTP Y AFTSSSML RRNTPLLGQA LTIASKHHQI VKMDLRSRYL DSLKAIVFKQ YNQTFETHNS TTPLQERSLG LDINMDSRII HE NIVEKER VQRITQETFG DYPQPQLEFA QYKLETKFKS GLNGSILAER EEPLRCLIKF SSPHLLEALK SLAPAGIADA PLS PLLTCI PNKRMNYFKI RDK

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Macromolecule #8: Centromere protein O

MacromoleculeName: Centromere protein O / type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 33.830637 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MEQANPLRPD GESKGGVLAH LERLETQVSR SRKQSEELQS VQAQEGALGT KIHKLRRLRD ELRAVVRHRR ASVKACIANV EPNQTVEIN EQEALEEKLE NVKAILQAYH FTGLSGKLTS RGVCVCISTA FEGNLLDSYF VDLVIQKPLR IHHHSVPVFI P LEEIAAKY ...String:
MEQANPLRPD GESKGGVLAH LERLETQVSR SRKQSEELQS VQAQEGALGT KIHKLRRLRD ELRAVVRHRR ASVKACIANV EPNQTVEIN EQEALEEKLE NVKAILQAYH FTGLSGKLTS RGVCVCISTA FEGNLLDSYF VDLVIQKPLR IHHHSVPVFI P LEEIAAKY LQTNIQHFLF SLCEYLNAYS GRKYQADRLQ SDFAALLTGP LQRNPLCNLL SFTYKLDPGG QSFPFCARLL YK DLTATLP TDVTVTCQGV EVLSTSWEEQ RASHETLFCT KPLHQVFASF TRKGEKLDMS LVS

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Macromolecule #9: Centromere protein P

MacromoleculeName: Centromere protein P / type: protein_or_peptide / ID: 9 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 33.210949 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MDAELAEVRA LQAEIAALRR ACEDPPAPWE EKSRVQKSFQ AIHQFNLEGW KSSKDLKNQL GHLESELSFL STLTGINIRN HSKQTEDLT STEMTEKSIR KVLQRHRLSG NCHMVTFQLE FQILEIQNKE RLSSAVTDLN IIMEPTECSE LSEFVSRAEE R KDLFMFFR ...String:
MDAELAEVRA LQAEIAALRR ACEDPPAPWE EKSRVQKSFQ AIHQFNLEGW KSSKDLKNQL GHLESELSFL STLTGINIRN HSKQTEDLT STEMTEKSIR KVLQRHRLSG NCHMVTFQLE FQILEIQNKE RLSSAVTDLN IIMEPTECSE LSEFVSRAEE R KDLFMFFR SLHFFVEWFE YRKRTFKHLK EKYPDAVYLS EGPSSCSMGI RSASRPGFEL VIVWRIQIDE DGKVFPKLDL LT KVPQRAL ELDKNRAIET APLSFRTLVG LLGIEAALES LIKSLCAEEN N

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Macromolecule #10: Centromere protein Q

MacromoleculeName: Centromere protein Q / type: protein_or_peptide / ID: 10 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 30.648375 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MSGKANASKK NAQQLKRNPK RKKDNEEVVL SENKVRNTVK KNKNHLKDLS SEGQTKHTNL KHGKTAASKR KTWQPLSKST RDHLQTMME SVIMTILSNS IKEKEEIQYH LNFLKKRLLQ QCETLKVPPK KMEDLTNVSS LLNMERARDK ANEEGLALLQ E EIDKMVET ...String:
MSGKANASKK NAQQLKRNPK RKKDNEEVVL SENKVRNTVK KNKNHLKDLS SEGQTKHTNL KHGKTAASKR KTWQPLSKST RDHLQTMME SVIMTILSNS IKEKEEIQYH LNFLKKRLLQ QCETLKVPPK KMEDLTNVSS LLNMERARDK ANEEGLALLQ E EIDKMVET TELMTGNIQS LKNKIQILAS EVEEEEERVK QMHQINSSGV LSLPELSQKT LKAPTLQKEI LALIPNQNAL LK DLDILHN SSQMKSMSTF IEEAYKKLDA S

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Macromolecule #11: Centromere protein U

MacromoleculeName: Centromere protein U / type: protein_or_peptide / ID: 11 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 47.609766 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MAPRGRRRPR PHRSEGARRS KNTLERTHSM KDKAGQKCKP IDVFDFPDNS DVSSIGRLGE NEKDEETYET FDPPLHSTAI YADEEEFSK HCGLSLSSTP PGKEAKRSSD TSGNEASEIE SVKISAKKPG RKLRPISDDS ESIEESDTRR KVKSAEKIST Q RHEVIRTT ...String:
MAPRGRRRPR PHRSEGARRS KNTLERTHSM KDKAGQKCKP IDVFDFPDNS DVSSIGRLGE NEKDEETYET FDPPLHSTAI YADEEEFSK HCGLSLSSTP PGKEAKRSSD TSGNEASEIE SVKISAKKPG RKLRPISDDS ESIEESDTRR KVKSAEKIST Q RHEVIRTT ASSELSEKPA ESVTSKKTGP LSAQPSVEKE NLAIESQSKT QKKGKISHDK RKKSRSKAIG SDTSDIVHIW CP EGMKTSD IKELNIVLPE FEKTHLEHQQ RIESKVCKAA IATFYVNVKE QFIKMLKESQ MLTNLKRKNA KMISDIEKKR QRM IEVQDE LLRLEPQLKQ LQTKYDELKE RKSSLRNAAY FLSNLKQLYQ DYSDVQAQEP NVKETYDSSS LPALLFKART LLGA ESHLR NINHQLEKLL DQG

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Macromolecule #12: Centromere protein R

MacromoleculeName: Centromere protein R / type: protein_or_peptide / ID: 12 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 20.228297 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString:
MPVKRSLKLD GLLEENSFDP SKITRKKSVI TYSPTTGTCQ MSLFASPTSS EEQKHRNGLS NEKRKKLNHP SLTESKESTT KDNDEFMML LSKVEKLSEE IMEIMQNLSS IQALEGSREL ENLIGISCAS HFLKREMQKT KELMTKVNKQ KLFEKSTGLP H KASRHLDS YEFLKAILN

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Macromolecule #13: Centromere protein T

MacromoleculeName: Centromere protein T / type: protein_or_peptide / ID: 13 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 60.502613 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MADHNPDSDS TPRTLLRRVL DTADPRTPRR PRSARAGARR ALLETASPRK LSGQTRTIAR GRSHGARSVG RSAHIQASGH LEEQTPRTL LKNILLTAPE SSILMPESVV KPVPAPQAVQ PSRQESSCGS LELQLPELEP PTTLAPGLLA PGRRKQRLRL S VFQQGVDQ ...String:
MADHNPDSDS TPRTLLRRVL DTADPRTPRR PRSARAGARR ALLETASPRK LSGQTRTIAR GRSHGARSVG RSAHIQASGH LEEQTPRTL LKNILLTAPE SSILMPESVV KPVPAPQAVQ PSRQESSCGS LELQLPELEP PTTLAPGLLA PGRRKQRLRL S VFQQGVDQ GLSLSQEPQG NADASSLTRS LNLTFATPLQ PQSVQRPGLA RRPPARRAVD VGAFLRDLRD TSLAPPNIVL ED TQPFSQP MVGSPNVYHS LPCTPHTGAE DAEQAAGRKT QSSGPGLQKN SPGKPAQFLA GEAEEVNAFA LGFLSTSSGV SGE DEVEPL HDGVEEAEKK MEEEGVSVSE MEATGAQGPS RVEEAEGHTE VTEAEGSQGT AEADGPGASS GDEDASGRAA SPES ASSTP ESLQARRHHQ FLEPAPAPGA AVLSSEPAEP LLVRHPPRPR TTGPRPRQDP HKAGLSHYVK LFSFYAKMPM ERKAL EMVE KCLDKYFQHL CDDLEVFAAH AGRKTVKPED LELLMRRQGL VTDQVSLHVL VERHLPLEYR QLLIPCAYSG NSVFPA Q

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Macromolecule #14: Centromere protein W

MacromoleculeName: Centromere protein W / type: protein_or_peptide / ID: 14 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 10.087236 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString:
MALSTIVSQR KQIKRKAPRG FLKRVFKRKK PQLRLEKSGD LLVHLNCLLF VHRLAEESRT NACASKCRVI NKEHVLAAAK VILKKSRG

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Macromolecule #15: Centromere protein S

MacromoleculeName: Centromere protein S / type: protein_or_peptide / ID: 15 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 15.917875 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString:
MEEEAETEEQ QRFSYQQRLK AAVHYTVGCL CEEVALDKEM QFSKQTIAAI SELTFRQCEN FAKDLEMFAR HAKRTTINTE DVKLLARRS NSLLKYITDK SEEIAQINLE RKAQKKKKSE DGSKNSRQPA EAGVVESEN

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Macromolecule #16: Centromere protein X

MacromoleculeName: Centromere protein X / type: protein_or_peptide / ID: 16 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 8.972415 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString:
MEGAGAGSGF RKELVSRLLH LHFKDDKTKV SGDALQLMVE LLKVFVVEAA VRGVRQAQAE DALRVDVDQL EKVLPQLLLD F

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Macromolecule #3: DNA (66-MER)

MacromoleculeName: DNA (66-MER) / type: dna / ID: 3 / Number of copies: 1 / Classification: DNA
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 16.459549 KDa
SequenceString: (DA)(DA)(DT)(DC)(DT)(DG)(DC)(DA)(DA)(DG) (DT)(DG)(DG)(DA)(DT)(DA)(DT)(DT)(DT)(DG) (DG)(DA)(DC)(DC)(DG)(DC)(DT)(DT)(DT) (DG)(DA)(DG)(DG)(DC)(DC)(DT)(DT)(DC)(DG) (DT) (DT)(DG)(DG)(DA)(DA)(DA) ...String:
(DA)(DA)(DT)(DC)(DT)(DG)(DC)(DA)(DA)(DG) (DT)(DG)(DG)(DA)(DT)(DA)(DT)(DT)(DT)(DG) (DG)(DA)(DC)(DC)(DG)(DC)(DT)(DT)(DT) (DG)(DA)(DG)(DG)(DC)(DC)(DT)(DT)(DC)(DG) (DT) (DT)(DG)(DG)(DA)(DA)(DA)(DC)(DG) (DG)(DG)(DA)(DA)(DT)

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Macromolecule #17: DNA (66-MER)

MacromoleculeName: DNA (66-MER) / type: dna / ID: 17 / Number of copies: 1 / Classification: DNA
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 16.197409 KDa
SequenceString: (DA)(DT)(DT)(DC)(DC)(DC)(DG)(DT)(DT)(DT) (DC)(DC)(DA)(DA)(DC)(DG)(DA)(DA)(DG)(DG) (DC)(DC)(DT)(DC)(DA)(DA)(DA)(DG)(DC) (DG)(DG)(DT)(DC)(DC)(DA)(DA)(DA)(DT)(DA) (DT) (DC)(DC)(DA)(DC)(DT)(DT) ...String:
(DA)(DT)(DT)(DC)(DC)(DC)(DG)(DT)(DT)(DT) (DC)(DC)(DA)(DA)(DC)(DG)(DA)(DA)(DG)(DG) (DC)(DC)(DT)(DC)(DA)(DA)(DA)(DG)(DC) (DG)(DG)(DT)(DC)(DC)(DA)(DA)(DA)(DT)(DA) (DT) (DC)(DC)(DA)(DC)(DT)(DT)(DG)(DC) (DA)(DG)(DA)(DT)(DT)

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Macromolecule #18: water

MacromoleculeName: water / type: ligand / ID: 18 / Number of copies: 1 / Formula: HOH
Molecular weightTheoretical: 18.015 Da
Chemical component information

ChemComp-HOH:
WATER / Water

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

BufferpH: 7.8
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Nominal defocus max: 3.0 µm / Nominal defocus min: 1.5 µm
Image recordingFilm or detector model: GATAN K3 (6k x 4k) / Average electron dose: 40.0 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Initial angle assignmentType: MAXIMUM LIKELIHOOD
Final angle assignmentType: MAXIMUM LIKELIHOOD
Final reconstructionResolution.type: BY AUTHOR / Resolution: 2.83 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 123677
FSC plot (resolution estimation)

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