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- EMDB-14098: Structure of the human inner kinetochore CCAN complex -

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Basic information

Entry
Database: EMDB / ID: EMD-14098
TitleStructure of the human inner kinetochore CCAN complex
Map datamain map
Sample
  • Complex: human inner kinetochore CCAN complex
    • Protein or peptide: x 15 types
Function / homology
Function and homology information


positive regulation of protein localization to kinetochore / Mis6-Sim4 complex / centromere complex assembly / kinetochore organization / metaphase chromosome alignment / spindle attachment to meiosis I kinetochore / inner kinetochore / kinetochore binding / centromeric DNA binding / sex differentiation ...positive regulation of protein localization to kinetochore / Mis6-Sim4 complex / centromere complex assembly / kinetochore organization / metaphase chromosome alignment / spindle attachment to meiosis I kinetochore / inner kinetochore / kinetochore binding / centromeric DNA binding / sex differentiation / CENP-A containing chromatin assembly / chordate embryonic development / negative regulation of epithelial cell apoptotic process / attachment of mitotic spindle microtubules to kinetochore / kinetochore assembly / condensed chromosome, centromeric region / mitotic sister chromatid segregation / chromosome, centromeric region / centriolar satellite / chromosome organization / Amplification of signal from unattached kinetochores via a MAD2 inhibitory signal / pericentric heterochromatin / 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 / chromosome segregation / positive regulation of epithelial cell proliferation / RHO GTPases Activate Formins / kinetochore / nuclear matrix / Separation of Sister Chromatids / actin cytoskeleton / mitotic cell cycle / chromosome / midbody / nuclear body / cell adhesion / protein heterodimerization activity / cell division / apoptotic process / nucleolus / regulation of DNA-templated transcription / signal transduction / DNA binding / nucleoplasm / membrane / identical protein binding / 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 / CENP-C, middle DNMT3B-binding domain / Centromere assembly component CENP-C middle DNMT3B-binding region / Centromere protein O / Cenp-O kinetochore centromere component / Kinetochore assembly subunit CENP-C, N-terminal domain / Kinetochore assembly subunit CENP-C N-terminal / Mif2/CENP-C cupin domain / Centromere protein C/Mif2/cnp3 / Mif2/CENP-C like / Centromere protein Chl4/mis15/CENP-N / Kinetochore protein CHL4 like / RmlC-like cupin domain superfamily / RmlC-like jelly roll fold / 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 C / Centromere protein R / Centromere protein W / Centromere protein P / Centromere protein U / Centromere protein Q / Centromere protein L / Centromere protein I / Centromere protein T / Centromere protein N ...Centromere protein C / Centromere protein R / Centromere protein W / Centromere protein P / Centromere protein U / Centromere protein Q / Centromere protein L / 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: 4.6 Å
AuthorsVetter IR / Pesenti M / Raisch T
Funding support1 items
OrganizationGrant numberCountry
Not funded
CitationJournal: Mol Cell / Year: 2022
Title: Structure of the human inner kinetochore CCAN complex and its significance for human centromere organization.
Authors: Marion E Pesenti / Tobias Raisch / Duccio Conti / Kai Walstein / Ingrid Hoffmann / Dorothee Vogt / Daniel Prumbaum / Ingrid R Vetter / Stefan Raunser / Andrea Musacchio /
Abstract: Centromeres are specialized chromosome loci that seed the kinetochore, a large protein complex that effects chromosome segregation. A 16-subunit complex, the constitutive centromere associated ...Centromeres are specialized chromosome loci that seed the kinetochore, a large protein complex that effects chromosome segregation. A 16-subunit complex, the constitutive centromere associated network (CCAN), connects between the specialized centromeric chromatin, marked by the histone H3 variant CENP-A, and the spindle-binding moiety of the kinetochore. Here, we report a cryo-electron microscopy structure of human CCAN. We highlight unique features such as the pseudo GTPase CENP-M and report how a crucial CENP-C motif binds the CENP-LN complex. The CCAN structure has implications for the mechanism of specific recognition of the CENP-A nucleosome. A model consistent with our structure depicts the CENP-C-bound nucleosome as connected to the CCAN through extended, flexible regions of CENP-C. An alternative model identifies both CENP-C and CENP-N as specificity determinants but requires CENP-N to bind CENP-A in a mode distinct from the classical nucleosome octamer.
History
DepositionDec 24, 2021-
Header (metadata) releaseJun 8, 2022-
Map releaseJun 8, 2022-
UpdateJun 15, 2022-
Current statusJun 15, 2022Processing site: PDBe / Status: Released

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Structure visualization

Supplemental images

Downloads & links

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Map

FileDownload / File: emd_14098.map.gz / Format: CCP4 / Size: 216 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Annotationmain map
Projections & slices

Image control

Size
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Others
AxesZ (Sec.)Y (Row.)X (Col.)
0.7 Å/pix.
x 384 pix.
= 268.8 Å
0.7 Å/pix.
x 384 pix.
= 268.8 Å
0.7 Å/pix.
x 384 pix.
= 268.8 Å

Surface

Projections

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Images are generated by Spider.

Voxel sizeX=Y=Z: 0.7 Å
Density
Contour LevelBy AUTHOR: 0.003
Minimum - Maximum-0.0010002586 - 0.020386236
Average (Standard dev.)0.00024873368 (±0.0006638026)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions384384384
Spacing384384384
CellA=B=C: 268.8 Å
α=β=γ: 90.0 °

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Supplemental data

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Mask #1

Fileemd_14098_msk_1.map
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Mask #2

Fileemd_14098_msk_2.map
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Mask #3

Fileemd_14098_msk_3.map
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Sample components

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Entire : human inner kinetochore CCAN complex

EntireName: human inner kinetochore CCAN complex
Components
  • Complex: human inner kinetochore CCAN complex
    • Protein or peptide: Centromere protein C
    • Protein or peptide: Centromere protein H
    • Protein or peptide: Centromere protein I
    • 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 U
    • Protein or peptide: Centromere protein Q
    • Protein or peptide: Centromere protein R
    • Protein or peptide: Centromere protein T
    • Protein or peptide: Centromere protein W
    • Protein or peptide: Unknown protein

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Supramolecule #1: human inner kinetochore CCAN complex

SupramoleculeName: human inner kinetochore CCAN complex / type: complex / Chimera: Yes / ID: 1 / Parent: 0 / Macromolecule list: all
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 451 KDa

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Macromolecule #1: Centromere protein C

MacromoleculeName: Centromere protein C / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 61.856004 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MAASGLDHLK NGYRRRFCRP SRARDINTEQ GQNVLEILQD CFEEKSLAND FSTNSTKSVP NSTRKIKDTC IQSPSKECQK SHPKSVPVS SKKKEASLQF VVEPSEATNR SVQAHEVHQK ILATDVSSKN TPDSKKISSR NINDHHSEAD EEFYLSVGSP S VLLDAKTS ...String:
MAASGLDHLK NGYRRRFCRP SRARDINTEQ GQNVLEILQD CFEEKSLAND FSTNSTKSVP NSTRKIKDTC IQSPSKECQK SHPKSVPVS SKKKEASLQF VVEPSEATNR SVQAHEVHQK ILATDVSSKN TPDSKKISSR NINDHHSEAD EEFYLSVGSP S VLLDAKTS VSQNVIPSSA QKRETYTFEN SVNMLPSSTE VSVKTKKRLN FDDKVMLKKI EIDNKVSDEE DKTSEGQERK PS GSSQNRI RDSEYEIQRQ AKKSFSTLFL ETVKRKSESS PIVRHAATAP PHSCPPDDTK LIEDEFIIDE SDQSFASRSW ITI PRKAGS LKQRTISPAE STALLQGRKS REKHHNILPK TLANDKHSHK PHPVETSQPS DKTVLDTSYA LIGETVNNYR STKY EMYSK NAEKPSRSKR TIKQKQRRKF MAKPAEEQLD VGQSKDENIH TSHITQDEFQ RNSDRNMEEH EEMGNDCVSK KQMPP VGSK KSSTRKDKEE SKKKRFSSES KNKLVPEEVT STVTKSRRIS RRPSDWWVVK SEESPVYSNS

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Macromolecule #2: Centromere protein H

MacromoleculeName: Centromere protein H / type: protein_or_peptide / ID: 2 / 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 #3: Centromere protein I

MacromoleculeName: Centromere protein I / type: protein_or_peptide / ID: 3 / 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 U

MacromoleculeName: Centromere protein U / type: protein_or_peptide / ID: 10 / 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 #11: Centromere protein Q

MacromoleculeName: Centromere protein Q / type: protein_or_peptide / ID: 11 / 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 #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: Escherichia coli BL21(DE3) (bacteria)
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: Escherichia coli BL21(DE3) (bacteria)
SequenceString:
MALSTIVSQR KQIKRKAPRG FLKRVFKRKK PQLRLEKSGD LLVHLNCLLF VHRLAEESRT NACASKCRVI NKEHVLAAAK VILKKSRG

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Macromolecule #15: Unknown protein

MacromoleculeName: Unknown protein / type: protein_or_peptide / ID: 15 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 1.294587 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString:
(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)

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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

Concentration1.5 mg/mL
BufferpH: 6.8 / Details: 0.0025% Triton
GridModel: UltrAuFoil R1.2/1.3 / Material: GOLD / Pretreatment - Type: GLOW DISCHARGE
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 286 K / Instrument: FEI VITROBOT MARK IV / Details: blot for 3.5 seconds at blot force -3.

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: OTHER / Imaging mode: BRIGHT FIELDBright-field microscopy / Nominal defocus max: 1.2 µm / Nominal defocus min: 0.6 µm / Nominal magnification: 130000
Specialist opticsPhase plate: VOLTA PHASE PLATE / Energy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
Image recordingFilm or detector model: GATAN K3 (6k x 4k) / Number real images: 1540 / Average electron dose: 76.8 e/Å2
Details: Images were collected in movie mode with 80 frames per image in superresolution mode with 0.35 A/px (0.7A native)
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Particle selectionNumber selected: 140910
CTF correctionSoftware - Name: CTFFIND
Startup modelType of model: INSILICO MODEL / In silico model: Alphafold2 predictions
Initial angle assignmentType: NOT APPLICABLE
Final 3D classificationNumber classes: 4 / Software - Name: RELION (ver. 3.1.2)
Final angle assignmentType: NOT APPLICABLE
Final reconstructionNumber classes used: 1 / Applied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 4.6 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 3.1.2) / Number images used: 25206
FSC plot (resolution estimation)

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Atomic model buiding 1

RefinementSpace: REAL / Protocol: AB INITIO MODEL
Output model

PDB-7qoo:
Structure of the human inner kinetochore CCAN complex

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