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- EMDB-14351: Structure of the human CCAN CENP-A alpha-satellite complex -

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Basic information

Entry
Database: EMDB / ID: EMD-14351
TitleStructure of the human CCAN CENP-A alpha-satellite complex
Map dataLocal resolution filtered cryosparc homogeneous refinement map
Sample
  • Complex: CCAN-CENP-A inner centromere complex
    • Protein or peptide: x 20 types
    • DNA: x 2 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 / spindle attachment to meiosis I kinetochore / Fanconi anaemia nuclear complex / inner kinetochore / kinetochore binding ...positive regulation of protein localization to kinetochore / Mis6-Sim4 complex / FANCM-MHF complex / centromere complex assembly / kinetochore organization / metaphase chromosome alignment / spindle attachment to meiosis I kinetochore / Fanconi anaemia nuclear complex / inner kinetochore / kinetochore binding / centromeric DNA binding / sex differentiation / CENP-A containing chromatin assembly / resolution of meiotic recombination intermediates / chordate embryonic development / protein localization to chromosome, centromeric region / negative regulation of epithelial cell apoptotic process / attachment of mitotic spindle microtubules to kinetochore / kinetochore assembly / condensed chromosome, centromeric region / replication fork processing / mitotic sister chromatid segregation / establishment of mitotic spindle orientation / mitotic cytokinesis / chromosome, centromeric region / centriolar satellite / chromosome organization / negative regulation of megakaryocyte differentiation / protein localization to CENP-A containing chromatin / Amplification of signal from unattached kinetochores via a MAD2 inhibitory signal / pericentric heterochromatin / Replacement of protamines by nucleosomes in the male pronucleus / interstrand cross-link repair / CENP-A containing nucleosome / Packaging Of Telomere Ends / Mitotic Prometaphase / EML4 and NUDC in mitotic spindle formation / Recognition and association of DNA glycosylase with site containing an affected purine / Cleavage of the damaged purine / Deposition of new CENPA-containing nucleosomes at the centromere / Resolution of Sister Chromatid Cohesion / Recognition and association of DNA glycosylase with site containing an affected pyrimidine / Cleavage of the damaged pyrimidine / Inhibition of DNA recombination at telomere / Meiotic synapsis / telomere organization / NRIF signals cell death from the nucleus / RNA Polymerase I Promoter Opening / SUMOylation of chromatin organization proteins / Assembly of the ORC complex at the origin of replication / mitotic spindle organization / DNA methylation / Condensation of Prophase Chromosomes / HCMV Late Events / Chromatin modifications during the maternal to zygotic transition (MZT) / ERCC6 (CSB) and EHMT2 (G9a) positively regulate rRNA expression / SIRT1 negatively regulates rRNA expression / innate immune response in mucosa / PRC2 methylates histones and DNA / positive regulation of protein ubiquitination / Defective pyroptosis / chromosome segregation / positive regulation of epithelial cell proliferation / RHO GTPases Activate Formins / HDACs deacetylate histones / Fanconi Anemia Pathway / RNA Polymerase I Promoter Escape / Nonhomologous End-Joining (NHEJ) / Transcriptional regulation by small RNAs / Formation of the beta-catenin:TCF transactivating complex / RUNX1 regulates genes involved in megakaryocyte differentiation and platelet function / PKR-mediated signaling / Activated PKN1 stimulates transcription of AR (androgen receptor) regulated genes KLK2 and KLK3 / NoRC negatively regulates rRNA expression / G2/M DNA damage checkpoint / B-WICH complex positively regulates rRNA expression / HDMs demethylate histones / DNA Damage/Telomere Stress Induced Senescence / Metalloprotease DUBs / kinetochore / PKMTs methylate histone lysines / RMTs methylate histone arginines / Meiotic recombination / Pre-NOTCH Transcription and Translation / nuclear matrix / nucleosome assembly / Activation of anterior HOX genes in hindbrain development during early embryogenesis / HCMV Early Events / Transcriptional regulation of granulopoiesis / structural constituent of chromatin / Separation of Sister Chromatids / UCH proteinases / nucleosome / antimicrobial humoral immune response mediated by antimicrobial peptide / actin cytoskeleton / E3 ubiquitin ligases ubiquitinate target proteins / mitotic cell cycle / Recruitment and ATM-mediated phosphorylation of repair and signaling proteins at DNA double strand breaks / chromosome / RUNX1 regulates transcription of genes involved in differentiation of HSCs
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 / 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 / Histone H2B signature. / Histone H2B / Histone H2B / Histone H2A conserved site / Histone H2A signature. / Histone H2A, C-terminal domain / C-terminus of histone H2A / Histone H2A / Histone 2A / Histone H4, conserved site / Histone H4 signature. / Histone H4 / Histone H4 / RmlC-like jelly roll fold / CENP-T/Histone H4, histone fold / Centromere kinetochore component CENP-T histone fold / TATA box binding protein associated factor / TATA box binding protein associated factor (TAF), histone-like fold domain / Histone H3 signature 2. / Histone H3 / Histone H3/CENP-A / Histone H2A/H2B/H3 / Core histone H2A/H2B/H3/H4 / Histone-fold / P-loop containing nucleoside triphosphate hydrolase
Similarity search - Domain/homology
Centromere protein X / Histone H3-like centromeric protein A / Histone H4 / Histone H2B type 1-C/E/F/G/I / Centromere protein C / Centromere protein R / Centromere protein W / Centromere protein P / Centromere protein U / Centromere protein Q ...Centromere protein X / Histone H3-like centromeric protein A / Histone H4 / Histone H2B type 1-C/E/F/G/I / Centromere protein C / Centromere protein R / Centromere protein W / Centromere protein P / Centromere protein U / Centromere protein Q / Centromere protein L / Centromere protein S / Centromere protein I / Histone H2A type 1-C / 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: 12.0 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 14, 2022-
Header (metadata) releaseMay 18, 2022-
Map releaseMay 18, 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_14351.map.gz / Format: CCP4 / Size: 18.7 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationLocal resolution filtered cryosparc homogeneous refinement map
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
1.72 Å/pix.
x 170 pix.
= 292.4 Å
1.72 Å/pix.
x 170 pix.
= 292.4 Å
1.72 Å/pix.
x 170 pix.
= 292.4 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 1.72 Å
Density
Contour LevelBy AUTHOR: 0.3
Minimum - Maximum-0.3765454 - 2.0383599
Average (Standard dev.)0.041697286 (±0.1794751)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions170170170
Spacing170170170
CellA=B=C: 292.4 Å
α=β=γ: 90.0 °

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Supplemental data

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Sample components

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Entire : CCAN-CENP-A inner centromere complex

EntireName: CCAN-CENP-A inner centromere complex
Components
  • Complex: CCAN-CENP-A inner centromere complex
    • 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 Q
    • Protein or peptide: Centromere protein U
    • Protein or peptide: Centromere protein P
    • 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 (171-MER)
    • DNA: DNA (171-MER)
    • Protein or peptide: Histone H3-like centromeric protein A
    • Protein or peptide: Histone H4
    • Protein or peptide: Histone H2A type 1-C
    • Protein or peptide: Histone H2B type 1-C/E/F/G/I
    • Protein or peptide: Centromere protein C

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Supramolecule #1: CCAN-CENP-A inner centromere complex

SupramoleculeName: CCAN-CENP-A inner centromere complex / type: complex / Chimera: Yes / ID: 1 / Parent: 0 / Macromolecule list: #1-#22

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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 #3: Centromere protein K

MacromoleculeName: Centromere protein K / type: protein_or_peptide / ID: 3 / 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 #4: Centromere protein L

MacromoleculeName: Centromere protein L / type: protein_or_peptide / ID: 4 / 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 #5: Centromere protein M

MacromoleculeName: Centromere protein M / type: protein_or_peptide / ID: 5 / 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 #6: Centromere protein N

MacromoleculeName: Centromere protein N / type: protein_or_peptide / ID: 6 / 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 #7: Centromere protein O

MacromoleculeName: Centromere protein O / type: protein_or_peptide / ID: 7 / 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 #8: Centromere protein Q

MacromoleculeName: Centromere protein Q / type: protein_or_peptide / ID: 8 / 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 #9: Centromere protein U

MacromoleculeName: Centromere protein U / type: protein_or_peptide / ID: 9 / 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 #10: Centromere protein P

MacromoleculeName: Centromere protein P / type: protein_or_peptide / ID: 10 / 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 #11: Centromere protein R

MacromoleculeName: Centromere protein R / type: protein_or_peptide / ID: 11 / 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 #12: Centromere protein T

MacromoleculeName: Centromere protein T / type: protein_or_peptide / ID: 12 / 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 #13: Centromere protein W

MacromoleculeName: Centromere protein W / type: protein_or_peptide / ID: 13 / 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 #14: Centromere protein S

MacromoleculeName: Centromere protein S / type: protein_or_peptide / ID: 14 / 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

+
Macromolecule #15: Centromere protein X

MacromoleculeName: Centromere protein X / type: protein_or_peptide / ID: 15 / 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

+
Macromolecule #18: Histone H3-like centromeric protein A

MacromoleculeName: Histone H3-like centromeric protein A / type: protein_or_peptide / ID: 18 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 16.02363 KDa
Recombinant expressionOrganism: Escherichia coli (E. coli)
SequenceString:
MGPRRRSRKP EAPRRRSPSP TPTPGPSRRG PSLGASSHQH SRRRQGWLKE IRKLQKSTHL LIRKLPFSRL AREICVKFTR GVDFNWQAQ ALLALQEAAE AFLVHLFEDA YLLTLHAGRV TLFPKDVQLA RRIRGLEEGL G

+
Macromolecule #19: Histone H4

MacromoleculeName: Histone H4 / type: protein_or_peptide / ID: 19 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 11.394426 KDa
Recombinant expressionOrganism: Escherichia coli (E. coli)
SequenceString:
MSGRGKGGKG LGKGGAKRHR KVLRDNIQGI TKPAIRRLAR RGGVKRISGL IYEETRGVLK VFLENVIRDA VTYTEHAKRK TVTAMDVVY ALKRQGRTLY GFGG

+
Macromolecule #20: Histone H2A type 1-C

MacromoleculeName: Histone H2A type 1-C / type: protein_or_peptide / ID: 20 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 14.135523 KDa
Recombinant expressionOrganism: Escherichia coli (E. coli)
SequenceString:
MSGRGKQGGK ARAKAKSRSS RAGLQFPVGR VHRLLRKGNY AERVGAGAPV YLAAVLEYLT AEILELAGNA ARDNKKTRII PRHLQLAIR NDEELNKLLG RVTIAQGGVL PNIQAVLLPK KTESHHKAKG K

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Macromolecule #21: Histone H2B type 1-C/E/F/G/I

MacromoleculeName: Histone H2B type 1-C/E/F/G/I / type: protein_or_peptide / ID: 21 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 13.937213 KDa
Recombinant expressionOrganism: Escherichia coli (E. coli)
SequenceString:
MPEPAKSAPA PKKGSKKAVT KAQKKDGKKR KRSRKESYSV YVYKVLKQVH PDTGISSKAM GIMNSFVNDI FERIAGEASR LAHYNKRST ITSREIQTAV RLLLPGELAK HAVSEGTKAV TKYTSSK

+
Macromolecule #22: Centromere protein C

MacromoleculeName: Centromere protein C / type: protein_or_peptide / ID: 22 / Number of copies: 2 / 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

+
Macromolecule #16: DNA (171-MER)

MacromoleculeName: DNA (171-MER) / type: dna / ID: 16 / Number of copies: 1 / Classification: DNA
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 52.725812 KDa
SequenceString: (DT)(DC)(DC)(DA)(DA)(DA)(DT)(DG)(DT)(DC) (DC)(DA)(DA)(DT)(DT)(DC)(DC)(DA)(DG)(DA) (DT)(DA)(DC)(DT)(DA)(DC)(DA)(DA)(DA) (DA)(DA)(DG)(DA)(DG)(DT)(DG)(DT)(DT)(DT) (DC) (DA)(DA)(DA)(DA)(DC)(DT) ...String:
(DT)(DC)(DC)(DA)(DA)(DA)(DT)(DG)(DT)(DC) (DC)(DA)(DA)(DT)(DT)(DC)(DC)(DA)(DG)(DA) (DT)(DA)(DC)(DT)(DA)(DC)(DA)(DA)(DA) (DA)(DA)(DG)(DA)(DG)(DT)(DG)(DT)(DT)(DT) (DC) (DA)(DA)(DA)(DA)(DC)(DT)(DG)(DC) (DT)(DC)(DT)(DA)(DT)(DG)(DA)(DA)(DA)(DA) (DG)(DG) (DA)(DA)(DT)(DG)(DT)(DT)(DC) (DA)(DA)(DC)(DT)(DC)(DT)(DA)(DT)(DG)(DA) (DG)(DT)(DT) (DG)(DA)(DA)(DT)(DG)(DC) (DA)(DA)(DA)(DC)(DA)(DT)(DC)(DA)(DC)(DA) (DT)(DA)(DG)(DA) (DA)(DG)(DT)(DT)(DT) (DC)(DT)(DG)(DA)(DG)(DA)(DA)(DT)(DG)(DC) (DT)(DT)(DC)(DT)(DG) (DT)(DC)(DT)(DA) (DG)(DT)(DT)(DT)(DT)(DT)(DA)(DT)(DG)(DT) (DG)(DA)(DA)(DC)(DA)(DT) (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)

+
Macromolecule #17: DNA (171-MER)

MacromoleculeName: DNA (171-MER) / type: dna / ID: 17 / Number of copies: 1 / Classification: DNA
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 52.822809 KDa
SequenceString: (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)(DA)(DT)(DG)(DT)(DT)(DC)(DA)(DC) (DA) (DT)(DA)(DA)(DA)(DA)(DA) ...String:
(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)(DA)(DT)(DG)(DT)(DT)(DC)(DA)(DC) (DA) (DT)(DA)(DA)(DA)(DA)(DA)(DC)(DT) (DA)(DG)(DA)(DC)(DA)(DG)(DA)(DA)(DG)(DC) (DA)(DT) (DT)(DC)(DT)(DC)(DA)(DG)(DA) (DA)(DA)(DC)(DT)(DT)(DC)(DT)(DA)(DT)(DG) (DT)(DG)(DA) (DT)(DG)(DT)(DT)(DT)(DG) (DC)(DA)(DT)(DT)(DC)(DA)(DA)(DC)(DT)(DC) (DA)(DT)(DA)(DG) (DA)(DG)(DT)(DT)(DG) (DA)(DA)(DC)(DA)(DT)(DT)(DC)(DC)(DT)(DT) (DT)(DT)(DC)(DA)(DT) (DA)(DG)(DA)(DG) (DC)(DA)(DG)(DT)(DT)(DT)(DT)(DG)(DA)(DA) (DA)(DC)(DA)(DC)(DT)(DC) (DT)(DT)(DT) (DT)(DT)(DG)(DT)(DA)(DG)(DT)(DA)(DT)(DC) (DT)(DG)(DG)(DA)(DA)(DT)(DT) (DG)(DG) (DA)(DC)(DA)(DT)(DT)(DT)(DG)(DG)(DA)

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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: 2.6 µm / Nominal defocus min: 1.2 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: 12.0 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 20549
FSC plot (resolution estimation)

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