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- EMDB-4464: Cryo-EM reconstruction of the APC/C-MCC-Cdk2-cyclinA2-Cks2 comple... -

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Basic information

Entry
Database: EMDB / ID: EMD-4464
TitleCryo-EM reconstruction of the APC/C-MCC-Cdk2-cyclinA2-Cks2 complex in the closed conformation
Map datasharpened map with B factor -30
Sample
  • Complex: Cryo-EM reconstruction of the APC/C-MCC-Cdk2-cyclinA2-Cks2 complex in the closed conformation
    • Protein or peptide: x 21 types
Biological speciesHomo sapiens (human)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.7 Å
AuthorsZhang S / Barford D
Funding support United Kingdom, 2 items
OrganizationGrant numberCountry
Medical Research Council (United Kingdom)MC_UP_1021/6 United Kingdom
Cancer Research UKC576/A14109 United Kingdom
CitationJournal: Nat Commun / Year: 2019
Title: Cyclin A2 degradation during the spindle assembly checkpoint requires multiple binding modes to the APC/C.
Authors: Suyang Zhang / Thomas Tischer / David Barford /
Abstract: The anaphase-promoting complex/cyclosome (APC/C) orchestrates cell cycle progression by controlling the temporal degradation of specific cell cycle regulators. Although cyclin A2 and cyclin B1 are ...The anaphase-promoting complex/cyclosome (APC/C) orchestrates cell cycle progression by controlling the temporal degradation of specific cell cycle regulators. Although cyclin A2 and cyclin B1 are both targeted for degradation by the APC/C, during the spindle assembly checkpoint (SAC), the mitotic checkpoint complex (MCC) represses APC/C's activity towards cyclin B1, but not cyclin A2. Through structural, biochemical and in vivo analysis, we identify a non-canonical D box (D2) that is critical for cyclin A2 ubiquitination in vitro and degradation in vivo. During the SAC, cyclin A2 is ubiquitinated by the repressed APC/C-MCC, mediated by the cooperative engagement of its KEN and D2 boxes, ABBA motif, and the cofactor Cks. Once the SAC is satisfied, cyclin A2 binds APC/C-Cdc20 through two mutually exclusive binding modes, resulting in differential ubiquitination efficiency. Our findings reveal that a single substrate can engage an E3 ligase through multiple binding modes, affecting its degradation timing and efficiency.
History
DepositionDec 11, 2018-
Header (metadata) releaseSep 11, 2019-
Map releaseSep 11, 2019-
UpdateNov 25, 2020-
Current statusNov 25, 2020Processing site: PDBe / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.035
  • Imaged by UCSF Chimera
  • Download
  • Surface view colored by radius
  • Surface level: 0.035
  • Imaged by UCSF Chimera
  • Download
Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

emd_4464.png

Downloads & links

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Map

FileDownload / File: emd_4464.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Annotationsharpened map with B factor -30
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
1.06 Å/pix.
x 400 pix.
= 424. Å		1.06 Å/pix.
x 400 pix.
= 424. Å		1.06 Å/pix.
x 400 pix.
= 424. Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 1.06 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.035 / Movie #1: 0.035
Minimum - Maximum-0.052832663 - 0.123928554
Average (Standard dev.)0.00046262454 (±0.00430203)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions400400400
Spacing400400400
CellA=B=C: 423.99997 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z1.061.061.06
M x/y/z400400400
origin x/y/z0.0000.0000.000
length x/y/z424.000424.000424.000
α/β/γ90.00090.00090.000
start NX/NY/NZ000
NX/NY/NZ320320320
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS400400400
D min/max/mean-0.0530.1240.000

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Supplemental data

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Half map: halfmap1

Fileemd_4464_half_map_1.map
Annotationhalfmap1
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: halfmap2

Fileemd_4464_half_map_2.map
Annotationhalfmap2
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Sample components

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Entire : Cryo-EM reconstruction of the APC/C-MCC-Cdk2-cyclinA2-Cks2 comple...

EntireName: Cryo-EM reconstruction of the APC/C-MCC-Cdk2-cyclinA2-Cks2 complex in the closed conformation
Components
  • Complex: Cryo-EM reconstruction of the APC/C-MCC-Cdk2-cyclinA2-Cks2 complex in the closed conformation
    • Protein or peptide: Apc1
    • Protein or peptide: Apc2
    • Protein or peptide: Apc3
    • Protein or peptide: Apc4
    • Protein or peptide: Apc5
    • Protein or peptide: Apc6
    • Protein or peptide: Apc7
    • Protein or peptide: Apc8
    • Protein or peptide: Apc10
    • Protein or peptide: Apc11
    • Protein or peptide: Apc12
    • Protein or peptide: Apc13
    • Protein or peptide: Apc15
    • Protein or peptide: Apc16
    • Protein or peptide: Cdc20
    • Protein or peptide: Cdk2
    • Protein or peptide: Cyclin A2
    • Protein or peptide: Cks2
    • Protein or peptide: BubR1
    • Protein or peptide: Mad2
    • Protein or peptide: Bub3

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Supramolecule #1: Cryo-EM reconstruction of the APC/C-MCC-Cdk2-cyclinA2-Cks2 comple...

SupramoleculeName: Cryo-EM reconstruction of the APC/C-MCC-Cdk2-cyclinA2-Cks2 complex in the closed conformation
type: complex / ID: 1 / Parent: 0 / Macromolecule list: all
Source (natural)Organism: Homo sapiens (human)
Recombinant expressionOrganism: unidentified baculovirus
Molecular weightTheoretical: 1.6 MDa

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Macromolecule #1: Apc1

MacromoleculeName: Apc1 / type: protein_or_peptide / ID: 1 / Enantiomer: LEVO
SequenceString: MSNFYEERTT MIAARDLQEF VPFGRDHCKH HPNALNLQLR QLQPASELWS SDGAAGLVGS LQEVTIHEK QKESWQLRKG VSEIGEDVDY DEELYVAGNM VIWSKGSKSQ AMSNFYEERT TMIAARDLQE FVPFGRDHCK HHPNALNLQL RQLQPASELW SSDGAAGLVG S ...String:
MSNFYEERTT MIAARDLQEF VPFGRDHCKH HPNALNLQLR QLQPASELWS SDGAAGLVGS LQEVTIHEK QKESWQLRKG VSEIGEDVDY DEELYVAGNM VIWSKGSKSQ AMSNFYEERT TMIAARDLQE FVPFGRDHCK HHPNALNLQL RQLQPASELW SSDGAAGLVG S LQEVTIHE KQKESWQLRK GVSEIGEDVD YDEELYVAGN MVIWSKGSKS QALAVYKAFT VD SPVQQAL WCDFIISQDK SEKAYSSNEV EKCICILQSS CINMHSIEGK DYIASLPFQV ANV WPTKYG LLFERSASSH EVPPGSPREP LPTMFSMLHP LDEITPLVCK SGSLFGSSRV QYVV DHAMK IVFLNTDPSI VMTYDAVQNV HSVWTLRRVK SEEENVVLKF SEQGGTPQNV ATSSS LTAH LRSLSKGDSP VTSPFQNYSS IHSQSRSTSS PSLHSRSPSI SNMAALSRAH SPALGV HSF SGVQRFNISS HNQSPKRHSI SHSPNSNSNG SFLAPETEPI VPELCIDHLW TETITNI RE KNSQASKVFI TSDLCGQKFL CFLVESQLQL RCVKFQESND KTQLIFGSVT NIPAKDAA P VEKIDTMLVL EGSGNLVLYT GVVRVGKVFI PGLPAPSLTM SNTMPRPSTP LDGVSTPKP LSKLLGSLDE VVLLSPVPEL RDSSKLHDSL YNEDCTFQQL GTYIHSIRDP VHNRVTLELS NGSMVRITI PEIATSELVQ TCLQAIKFIL PKEIAVQMLV KWYNVHSAPG GPSYHSEWNL F VTCLMNMM GYNTDRLAWT RNFDFEGSLS PVIAPKKARP SETGSDDDWE YLLNSDYHQN VE SHLLNRS LCLSPSEASQ MKDEDFSQNL SLDSSTLLFT HIPAIFFVLH LVYEELKLNT LMG EGICSL VELLVQLARD LKLGPYVDHY YRDYPTLVRT TGQVCTIDPG QTGFMHHPSF FTSE PPSIY QWVSSCLKGE GMPPYPYLPG ICERSRLVVL SIALYILGDE SLVSDESSQY LTRIT IAPQ KLQVEQEENR FSFRHSTSVS SLAERLVVWM TNVGFTLRDL ETLPFGIALP IRDAIY HCR EQPASDWPEA VCLLIGRQDL SKQACEGNLP KGKSVLSSDV PSGTETEEED DGMNDMN HE VMSLIWSEDL RVQDVRRLLQ SAHPVRVNVV QYPELSDHEF IEEKENRLLQ LCQRTMAL P VGRGMFTLFS YHPVPTEPLP IPKLNLTGRA PPRNTTVDLN SGNIDVPPNM TSWASFHNG VAAGLKIAPA SQIDSAWIVY NKPKHAELAN EYAGFLMALG LNGHLTKLAT LNIHDYLTKG HEMTSIGLL LGVSAAKLGT MDMSITRLLS IHIPALLPPT STELDVPHNV QVAAVVGIGL V YQGTAHRH TAEVLLAEIG RPPGPEMEYC TDRESYSLAA GLALGMVCLG HGSNLIGMSD LN VPEQLYQ YMVGGHRRFQ TGMHREKHKS PSYQIKEGDT INVDVTCPGA TLALAMIYLK TNN RSIADW LRAPDTMYLL DFVKPEFLLL RTLARCLILW DDILPNSKWV DSNVPQIIRE NSIS LSEIE LPCSEDLNLE TLSQAHVYII AGACLSLGFR FAGSENLSAF NCLHKFAKDF MTYLS APNA SVTGPHNLET CLSVVLLSLA MVMAGSGNLK VLQLCRFLHM KTGGEMNYGF HLAHHM ALG LLFLGGGRYS LSTSNSSIAA LLCALYPHFP AHSTDNRYHL QALRHLYVLA AEPRLLV PV DVDTNTPCYA LLEVTYKGTQ WYEQTKEELM APTLLPELHL LKQIKVKGPR YWELLIDL S KGTQHLKSIL SKDGVLYVKL RAGQLSYKED PMGWQSLLAQ TVANRNSEAR AFKPETISA FTSDPALLSF AEYFCKPTVN MGQKQEILDL FSSVLYECVT QETPEMLPAY IAMDQAIRRL GRREMSETS ELWQIKLVLE FFSSRSHQER LQNHPKRGLF MNSEFLPVVK CTIDNTLDQW L QVGGDMCV HAYLSGQPLE ESQLSMLACF LVYHSVPAPQ HLPPIGLEGS TSFAELLFKF KQ LKMPVRA LLRLAPLLLG NPQPMVM LA VYKAFTVD S PVQQALWCDF IISQDKSEKA YSSNEVEKCI CILQSSCINM HSIEGKDYIA SLPFQVANV WPTKYGLLFE RSASSHEVPP GSPREPLPTM FSMLHPLDEI TPLVCKSGSL FGSSRVQYVV DHAMKIVFL NTDPSIVMTY DAVQNVHSVW TLRRVKSEEE NVVLKFSEQG GTPQNVATSS S LTAHLRSL SKGDSPVTSP FQNYSSIHSQ SRSTSSPSLH SRSPSISNMA ALSRAHSPAL GV HSFSGVQ RFNISSHNQS PKRHSISHSP NSNSNGSFLA PETEPIVPEL CIDHLWTETI TNI REKNSQ ASKVFITSDL CGQKFLCFLV ESQLQLRCVK FQESNDKTQL IFGSVTNIPA KDAA PVEKI DTMLVLEGSG NLVLYTGVVR VGKVFIPGLP APSLTMSNTM PRPSTPLDGV STPKP LSKL LGSLDEVVLL SPVPELRDSS KLHDSLYNED CTFQQLGTYI HSIRDPVHNR VTLELS NGS MVRITIPEIA TSELVQTCLQ AIKFILPKEI AVQMLVKWYN VHSAPGGPSY HSEWNLF VT CLMNMMGYNT DRLAWTRNFD FEGSLSPVIA PKKARPSETG SDDDWEYLLN SDYHQNVE S HLLNRSLCLS PSEASQMKDE DFSQNLSLDS STLLFTHIPA IFFVLHLVYE ELKLNTLMG EGICSLVELL VQLARDLKLG PYVDHYYRDY PTLVRTTGQV CTIDPGQTGF MHHPSFFTSE PPSIYQWVS SCLKGEGMPP YPYLPGICER SRLVVLSIAL YILGDESLVS DESSQYLTRI T IAPQKLQV EQEENRFSFR HSTSVSSLAE RLVVWMTNVG FTLRDLETLP FGIALPIRDA IY HCREQPA SDWPEAVCLL IGRQDLSKQA CEGNLPKGKS VLSSDVPSGT ETEEEDDGMN DMN HEVMSL IWSEDLRVQD VRRLLQSAHP VRVNVVQYPE LSDHEFIEEK ENRLLQLCQR TMAL PVGRG MFTLFSYHPV PTEPLPIPKL NLTGRAPPRN TTVDLNSGNI DVPPNMTSWA SFHNG VAAG LKIAPASQID SAWIVYNKPK HAELANEYAG FLMALGLNGH LTKLATLNIH DYLTKG HEM TSIGLLLGVS AAKLGTMDMS ITRLLSIHIP ALLPPTSTEL DVPHNVQVAA VVGIGLV YQ GTAHRHTAEV LLAEIGRPPG PEMEYCTDRE SYSLAAGLAL GMVCLGHGSN LIGMSDLN V PEQLYQYMVG GHRRFQTGMH REKHKSPSYQ IKEGDTINVD VTCPGATLAL AMIYLKTNN RSIADWLRAP DTMYLLDFVK PEFLLLRTLA RCLILWDDIL PNSKWVDSNV PQIIRENSIS LSEIELPCS EDLNLETLSQ AHVYIIAGAC LSLGFRFAGS ENLSAFNCLH KFAKDFMTYL S APNASVTG PHNLETCLSV VLLSLAMVMA GSGNLKVLQL CRFLHMKTGG EMNYGFHLAH HM ALGLLFL GGGRYSLSTS NSSIAALLCA LYPHFPAHST DNRYHLQALR HLYVLAAEPR LLV PVDVDT NTPCYALLEV TYKGTQWYEQ TKEELMAPTL LPELHLLKQI KVKGPRYWEL LIDL SKGTQ HLKSILSKDG VLYVKLRAGQ LSYKEDPMGW QSLLAQTVAN RNSEARAFKP ETISA FTSD PALLSFAEYF CKPTVNMGQK QEILDLFSSV LYECVTQETP EMLPAYIAMD QAIRRL GRR EMSETSELWQ IKLVLEFFSS RSHQERLQNH PKRGLFMNSE FLPVVKCTID NTLDQWL QV GGDMCVHAYL SGQPLEESQL SMLACFLVYH SVPAPQHLPP IGLEGSTSFA ELLFKFKQ L KMPVRALLRL APLLLGNPQP MVM

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Macromolecule #2: Apc2

MacromoleculeName: Apc2 / type: protein_or_peptide / ID: 2 / Enantiomer: LEVO
SequenceString: MAAAVVVAEG DSDSRPGQEL LVAWNTVSTG LVPPAALGLV SSRTSGAVPP KEEELRAAVE VLRGHGLHSV LEEWFVEVLQ NDLQANISP EFWNAISQCE NSADEPQCLL LLLDAFGLLE SRLDPYLRSL ELLEKWTRLG LLMGTGAQGL REEVHTMLRG V LFFSTPRT ...String:
MAAAVVVAEG DSDSRPGQEL LVAWNTVSTG LVPPAALGLV SSRTSGAVPP KEEELRAAVE VLRGHGLHSV LEEWFVEVLQ NDLQANISP EFWNAISQCE NSADEPQCLL LLLDAFGLLE SRLDPYLRSL ELLEKWTRLG LLMGTGAQGL REEVHTMLRG V LFFSTPRT FQEMIQRLYG CFLRVYMQSK RKGEGGTDPE LEGELDSRYA RRRYYRLLQS PLCAGCSSDK QQCWCRQALE QF HQLSQVL HRLSLLERVS AEAVTTTLHQ VTRERMEDRC RGEYERSFLR EFHKWIERVV GWLGKVFLQD GPARPASPEA GNT LRRWRC HVQRFFYRIY ASLRIEELFS IVRDFPDSRP AIEDLKYCLE RTDQRQQLLV SLKAALETRL LHPGVNTCDI ITLY ISAIK ALRVLDPSMV ILEVACEPIR RYLRTREDTV RQIVAGLTGD SDGTGDLAVE LSKTDPASLE TGQDSEDDSG EPEDW VPDP VDADPGKSSS KRRSSDIISL LVSIYGSKDL FINEYRSLLA DRLLHQFSFS PEREIRNVEL LKLRFGEAPM HFCEVM LKD MADSRRINAN IREEDEKRPA EEQPPFGVYA VILSSEFWPP FKDEKLEVPE DIRAALEAYC KKYEQLKAMR TLSWKHT LG LVTMDVELAD RTLSVAVTPV QAVILLYFQD QASWTLEELS KAVKMPVALL RRRMSVWLQQ GVLREEPPGT FSVIEEER P QDRDNMVLID SDDESDSGMA SQADQKEEEL LLFWTYIQAM LTNLESLSLD RIYNMLRMFV VTGPALAEID LQELQGYLQ KKVRDQQLVY SAGVYRLPKN CS

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Macromolecule #3: Apc3

MacromoleculeName: Apc3 / type: protein_or_peptide / ID: 3 / Enantiomer: LEVO
SequenceString: MTVLQEPVQA AIWQALNHYA YRDAVFLAER LYAEVHSEEA LFLLATCYYR SGKAYKAYRL LKGHSCTTPQ CKYLLAKCCV DLSKLAEGE QILSGGVFNK QKSHDDIVTE FGDSACFTLS LLGHVYCKTD RLAKGSECYQ KSLSLNPFLW SPFESLCEIG E KPDPDQTF ...String:
MTVLQEPVQA AIWQALNHYA YRDAVFLAER LYAEVHSEEA LFLLATCYYR SGKAYKAYRL LKGHSCTTPQ CKYLLAKCCV DLSKLAEGE QILSGGVFNK QKSHDDIVTE FGDSACFTLS LLGHVYCKTD RLAKGSECYQ KSLSLNPFLW SPFESLCEIG E KPDPDQTF KFTSLQNFSN CLPNSCTTQV PNHSLSHRQP ETVLTETPQD TIELNRLNLE SSNSKYSLNT DSSVSYIDSA VI SPDTVPL GTGTSILSKQ VQNKPKTGRS LLGGPAALSP LTPSFGILPL ETPSPGDGSY LQNYTNTPPV IDVPSTGAPS KKS VARIGQ TGTKSVFSQS GNSREVTPIL AQTQSSGPQT STTPQVLSPT ITSPPNALPR RSSRLFTSDS STTKENSKKL KMKF PPKIP NRKTKSKTNK GGITQPNIND SLEITKLDSS IISEGKISTI TPQIQAFNLQ KAAAEGLMSL LREMGKGYLA LCSYN CKEA INILSHLPSH HYNTGWVLCQ IGRAYFELSE YMQAERIFSE VRRIENYRVE GMEIYSTTLW HLQKDVALSV LSKDLT DMD KNSPEAWCAA GNCFSLQREH DIAIKFFQRA IQVDPNYAYA YTLLGHEFVL TEELDKALAC FRNAIRVNPR HYNAWYG LG MIYYKQEKFS LAEMHFQKAL DINPQSSVLL CHIGVVQHAL KKSEKALDTL NKAIVIDPKN PLCKFHRASV LFANEKYK S ALQELEELKQ IVPKESLVYF LIGKVYKKLG QTHLALMNFS WAMDLDPKGA NNQIKEAIDK RYLPDDEEPI TQEEQIMGT DESQESSMTD ADDTQLHAAE SDEF

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Macromolecule #4: Apc4

MacromoleculeName: Apc4 / type: protein_or_peptide / ID: 4 / Enantiomer: LEVO
SequenceString: MLRFPTCFPS FRVVGEKQLP QEIIFLVWSP KRDLIALANT AGEVLLHRLA SFHRVWSFPP NENTGKEVTC LAWRPDGKLL AFALADTKK IVLCDVEKPE SLHSFSVEAP VSCMHWMEVT VESSVLTSFY NAEDESNLLL PKLPTLPKNY SNTSKIFSEE N SDEIIKLL ...String:
MLRFPTCFPS FRVVGEKQLP QEIIFLVWSP KRDLIALANT AGEVLLHRLA SFHRVWSFPP NENTGKEVTC LAWRPDGKLL AFALADTKK IVLCDVEKPE SLHSFSVEAP VSCMHWMEVT VESSVLTSFY NAEDESNLLL PKLPTLPKNY SNTSKIFSEE N SDEIIKLL GDVRLNILVL GGSSGFIELY AYGMFKIARV TGIAGTCLAL CLSSDLKSLS VVTEVSTNGA SEVSYFQLET NL LYSFLPE VTRMARKFTH ISALLQYINL SLTCMCEAWE EILMQMDSRL TKFVQEKNTT TSVQDEFMHL LLWGKASAEL QTL LMNQLT VKGLKKLGQS IESSYSSIQK LVISHLQSGS ESLLYHLSEL KGMASWKQKY EPLGLDAAGI EEAITAVGSF ILKA NELLQ VIDSSMKNFK AFFRWLYVAM LRMTEDHVLP ELNKMTQKDI TFVAEFLTEH FNEAPDLYNR KGKYFNVERV GQYLK DEDD DLVSPPNTEG NQWYDFLQNS SHLKESPLLF PYYPRKSLHF VKRRMENIID QCLQKPADVI GKSMNQAICI PLYRDT RSE DSTRRLFKFP FLWNNKTSNL HYLLFTILED SLYKMCILRR HTDISQSVSN GLIAIKFGSF TYATTEKVRR SIYSCLD AQ FYDDETVTVV LKDTVGREGR DRLLVQLPLS LVYNSEDSAE YQFTGTYSTR LDEQCSAIPT RTMHFEKHWR LLESMKAQ Y VAGNGFRKVS CVLSSNLRHV RVFEMDIDDE WELDESSDEE EEASNKPVKI KEEVLSESEA ENQQAGAAAL APEIVIKVE KLDPELDS

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Macromolecule #5: Apc5

MacromoleculeName: Apc5 / type: protein_or_peptide / ID: 5 / Enantiomer: LEVO
SequenceString: MASVHESLYF NPMMTNGVVH ANVFGIKDWV TPYKIAVLVL LNEMSRTGEG AVSLMERRRL NQLLLPLLQ GPDITLSKLY KLIEESCPQL ANSVQIRIKL MAEGELKDME QFFDDLSDSF S GTEPEVHK TSVVGLFLRH MILAYSKLSF SQVFKLYTAL QQYFQNGEKK ...String:
MASVHESLYF NPMMTNGVVH ANVFGIKDWV TPYKIAVLVL LNEMSRTGEG AVSLMERRRL NQLLLPLLQ GPDITLSKLY KLIEESCPQL ANSVQIRIKL MAEGELKDME QFFDDLSDSF S GTEPEVHK TSVVGLFLRH MILAYSKLSF SQVFKLYTAL QQYFQNGEKK TVEDADMELT SR DEGERKM EKEELDVSVR EEEVSCSGPL SQKQAEFFLS QQASLLKNDE TKALTPASLQ KEL NNLLKF NPDFAEAHYL SYLNNLRVQD VFSSTHSLLH YFDRLILTGA ESKSNGEEGY GRSL RYAAL NLAALHCRFG HYQQAELALQ EAIRIAQESN DHVCLQHCLS WLYVLGQKRS DSYVL LEHS VKKAVHFGLP YLASLGIQSL VQQRAFAGKT ANKLMDALKD SDLLHWKHSL SELIDI SIA QKTAIWRLYG RSTMALQQAQ MLLSMNSLEA VNAGVQQNNT ESFAVALCHL AELHAEQ GC FAAASEVLKH LKERFPPNSQ HAQLWMLCDQ KIQFDRAMND GKYHLADSLV TGITALNS I EGVYRKAVVL QAQNQMSEAH KLLQKLLVHC QKLKNTEMVI SVLLSVAELY WRSSSPTIA LPMLLQALAL SKEYRLQYLA SETVLNLAFA QLILGIPEQA LSLLHMAIEP ILADGAILDK GRAMFLVAK CQVASAASYD QPKKAEALEA AIENLNEAKN YFAKVDCKER IRDVVYFQAR L YHTLGKTQ ERNRCAMLFR QLHQELPSHG VPLINHL

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Macromolecule #6: Apc6

MacromoleculeName: Apc6 / type: protein_or_peptide / ID: 6 / Enantiomer: LEVO
SequenceString: MNLERLRKRV RQYLDQQQYQ SALFWADKVA SLSREEPQDI YWLAQCLYLT AQYHRAAHAL RSRKLDKLY EACRYLAARC HYAAKEHQQA LDVLDMEEPI NKRLFEKYLK DESGFKDPSS D WEMSQSSI KSSICLLRGK IYDALDNRTL ATYSYKEALK LDVYCFEAFD ...String:
MNLERLRKRV RQYLDQQQYQ SALFWADKVA SLSREEPQDI YWLAQCLYLT AQYHRAAHAL RSRKLDKLY EACRYLAARC HYAAKEHQQA LDVLDMEEPI NKRLFEKYLK DESGFKDPSS D WEMSQSSI KSSICLLRGK IYDALDNRTL ATYSYKEALK LDVYCFEAFD LLTSHHMLTA QE EKELLES LPLSKLCNEE QELLRFLFEN KLKKYNKPSE TVIPESVDGL QENLDVVVSL AER HYYNCD FKMCYKLTSV VMEKDPFHAS CLPVHIGTLV ELNKANELFY LSHKLVDLYP SNPV SWFAV GCYYLMVGHK NEHARRYLSK ATTLEKTYGP AWIAYGHSFA VESEHDQAMA AYFTA AQLM KGCHLPMLYI GLEYGLTNNS KLAERFFSQA LSIAPEDPFV MHEVGVVAFQ NGEWKT AEK WFLDALEKIK AIGNEVTVDK WEPLLNNLGH VCRKLKKYAE ALDYHRQALV LIPQNAS TY SAIGYIHSLM GNFENAVDYF HTALGLRRDD TFSVTMLGHC IEMYIGDSEA YIGADIKD K LKCYDFDVHT MKTLKNIISP PWDFREFEVE KQTAEETGLT PLETSRKTPD SRPSLEETF EIEMNESDMM LETSMSDHST

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Macromolecule #7: Apc7

MacromoleculeName: Apc7 / type: protein_or_peptide / ID: 7 / Enantiomer: LEVO
SequenceString: MDPGDAAILE SSLRILYRLF ESVLPPLPAA LQSRMNVIDH VRDMAAAGLH SNVRLLSSLL LTMSNNNPE LFSPPQKYQL LVYHADSLFH DKEYRNAVSK YTMALQQKKA LSKTSKVRPS T GNSASTPQ SQCLPSEIEV KYKMAECYTM LKQDKDAIAI LDGIPSRQRT ...String:
MDPGDAAILE SSLRILYRLF ESVLPPLPAA LQSRMNVIDH VRDMAAAGLH SNVRLLSSLL LTMSNNNPE LFSPPQKYQL LVYHADSLFH DKEYRNAVSK YTMALQQKKA LSKTSKVRPS T GNSASTPQ SQCLPSEIEV KYKMAECYTM LKQDKDAIAI LDGIPSRQRT PKINMMLANL YK KAGQERP SVTSYKEVLR QCPLALDAIL GLLSLSVKGA EVASMTMNVI QTVPNLDWLS VWI KAYAFV HTGDNSRAIS TICSLEKKSL LRDNVDLLGS LADLYFRAGD NKNSVLKFEQ AQML DPYLI KGMDVYGYLL AREGRLEDVE NLGCRLFNIS DQHAEPWVVS GCHSFYSKRY SRALY LGAK AIQLNSNSVQ ALLLKGAALR NMGRVQEAII HFREAIRLAP CRLDCYEGLI ECYLAS NSI REAMVMANNV YKTLGANAQT LTLLATVCLE DPVTQEKAKT LLDKALTQRP DYIKAVV KK AELLSREQKY EDGIALLRNA LANQSDCVLH RILGDFLVAV NEYQEAMDQY SIALSLDP N DQKSLEGMQK MEKEESPTDA TQEEDVDDME GSGEEGDLEG SDSEAAQWAD QEQWFGMQ

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Macromolecule #8: Apc8

MacromoleculeName: Apc8 / type: protein_or_peptide / ID: 8 / Enantiomer: LEVO
SequenceString: MAASTSMVPV AVTAAVAPVL SINSDFSDLR EIKKQLLLIA GLTRERGLLH SSKWSAELAF SLPALPLAE LQPPPPITEE DAQDMDAYTL AKAYFDVKEY DRAAHFLHGC NSKKAYFLYM Y SRYLSGEK KKDDETVDSL GPLEKGQVKN EALRELRVEL SKKHQARELD ...String:
MAASTSMVPV AVTAAVAPVL SINSDFSDLR EIKKQLLLIA GLTRERGLLH SSKWSAELAF SLPALPLAE LQPPPPITEE DAQDMDAYTL AKAYFDVKEY DRAAHFLHGC NSKKAYFLYM Y SRYLSGEK KKDDETVDSL GPLEKGQVKN EALRELRVEL SKKHQARELD GFGLYLYGVV LR KLDLVKE AIDVFVEATH VLPLHWGAWL ELCNLITDKE MLKFLSLPDT WMKEFFLAHI YTE LQLIEE ALQKYQNLID VGFSKSSYIV SQIAVAYHNI RDIDKALSIF NELRKQDPYR IENM DTFSN LLYVRSMKSE LSYLAHNLCE IDKYRVETCC VIGNYYSLRS QHEKAALYFQ RALKL NPRY LGAWTLMGHE YMEMKNTSAA IQAYRHAIEV NKRDYRAWYG LGQTYEILKM PFYCLY YYR RAHQLRPNDS RMLVALGECY EKLNQLVEAK KCYWRAYAVG DVEKMALVKL AKLHEQL TE SEQAAQCYIK YIQDIYSCGE IVEHLEESTA FRYLAQYYFK CKLWDEASTC AQKCCAFN D TREEGKALLR QILQLRNQGE TPTTEVPAPF FLPASLSANN TPTRRVSPLN LSSVTP

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Macromolecule #9: Apc10

MacromoleculeName: Apc10 / type: protein_or_peptide / ID: 9 / Enantiomer: LEVO
SequenceString:
MTTPNKTPPG ADPKQLERTG TVREIGSQAV WSLSSCKPGF GVDQLRDDNL ETYWQSDGSQ PHLVNIQFR RKTTVKTLCI YADYKSDESY TPSKISVRVG NNFHNLQEIR QLELVEPSGW I HVPLTDNH KKPTRTFMIQ IAVLANHQNG RDTHMRQIKI YTPVEESSIG KFPRCTTIDF MM YRSIR

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Macromolecule #10: Apc11

MacromoleculeName: Apc11 / type: protein_or_peptide / ID: 10 / Enantiomer: LEVO
SequenceString:
MKVKIKCWNG VATWLWVAND ENCGICRMAF NGCCPDCKVP GDDCPLVWGQ CSHCFHMHCI LKWLHAQQV QQHCPMCRQE WKFKE

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Macromolecule #11: Apc12

MacromoleculeName: Apc12 / type: protein_or_peptide / ID: 11 / Enantiomer: LEVO
SequenceString:
MLRRKPTRLE LKLDDIEEFE NIRKDLETRK KQKEDVEVVG GSDGEGAIGL SSDPKSREQM INDRIGYKP QPKPNNRSSQ FGSLEF

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Macromolecule #12: Apc13

MacromoleculeName: Apc13 / type: protein_or_peptide / ID: 12 / Enantiomer: LEVO
SequenceString:
MDSEVQRDGR ILDLIDDAWR EDKLPYEDVA IPLNELPEPE QDNGGTTESV KEQEMKWTDL ALQYLHENV PPIGN

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Macromolecule #13: Apc15

MacromoleculeName: Apc15 / type: protein_or_peptide / ID: 13 / Enantiomer: LEVO
SequenceString:
MSTLFPSLFP RVTETLWFNL DRPCVEETEL QQQEQQHQAW LQSIAEKDNN LVPIGKPASE HYDDEEEED DEDDEDSEED SEDDEDMQDM DEMNDYNESP DDGEVNEVDM EGNEQDQDQW M I

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Macromolecule #14: Apc16

MacromoleculeName: Apc16 / type: protein_or_peptide / ID: 14 / Enantiomer: LEVO
SequenceString:
MAASSSSSSA GGVSGSSVTG SGFSVSDLAP PRKALFTYPK GAGEMLEDGS ERFLCESVFS YQVASTLKQ VKHDQQVARM EKLAGLVEEL EADEWRFKPI EQLLGFTPSS G

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Macromolecule #15: Cdc20

MacromoleculeName: Cdc20 / type: protein_or_peptide / ID: 15 / Enantiomer: LEVO
SequenceString: MAQFAFESDL HSLLQLDAPI PNAPPARWQR KAKEAAGPAP SPMRAANRSH SAGRTPGRTP GKSSSKVQT TPSKPGGDRY IPHRSAAQME VASFLLSKEN QPENSQTPTK KEHQKAWALN L NGFDVEEA KILRLSGKPQ NAPEGYQNRL KVLYSQKATP GSSRKTCRYI ...String:
MAQFAFESDL HSLLQLDAPI PNAPPARWQR KAKEAAGPAP SPMRAANRSH SAGRTPGRTP GKSSSKVQT TPSKPGGDRY IPHRSAAQME VASFLLSKEN QPENSQTPTK KEHQKAWALN L NGFDVEEA KILRLSGKPQ NAPEGYQNRL KVLYSQKATP GSSRKTCRYI PSLPDRILDA PE IRNDYYL NLVDWSSGNV LAVALDNSVY LWSASSGDIL QLLQMEQPGE YISSVAWIKE GNY LAVGTS SAEVQLWDVQ QQKRLRNMTS HSARVGSLSW NSYILSSGSR SGHIHHHDVR VAEH HVATL SGHSQEVCGL RWAPDGRHLA SGGNDNLVNV WPSAPGEGGW VPLQTFTQHQ GAVKA VAWC PWQSNVLATG GGTSDRHIRI WNVCSGACLS AVDAHSQVCS ILWSPHYKEL ISGHGF AQN QLVIWKYPTM AKVAELKGHT SRVLSLTMSP DGATVASAAA DETLRLWRCF ELDPARR RE REKASAAKSS LIHQGIR

+
Macromolecule #16: Cdk2

MacromoleculeName: Cdk2 / type: protein_or_peptide / ID: 16 / Enantiomer: LEVO
SequenceString: MENFQKVEKI GEGTYGVVYK ARNKLTGEVV ALKKIRLDTE TEGVPSTAIR EISLLKELNH PNIVKLLDV IHTENKLYLV FEFLHQDLKK FMDASALTGI PLPLIKSYLF QLLQGLAFCH S HRVLHRDL KPQNLLINTE GAIKLADFGL ARAFGVPVRT YTHEVVTLWY ...String:
MENFQKVEKI GEGTYGVVYK ARNKLTGEVV ALKKIRLDTE TEGVPSTAIR EISLLKELNH PNIVKLLDV IHTENKLYLV FEFLHQDLKK FMDASALTGI PLPLIKSYLF QLLQGLAFCH S HRVLHRDL KPQNLLINTE GAIKLADFGL ARAFGVPVRT YTHEVVTLWY RAPEILLGCK YY STAVDIW SLGCIFAEMV TRRALFPGDS EIDQLFRIFR TLGTPDEVVW PGVTSMPDYK PSF PKWARQ DFSKVVPPLD EDGRSLLSQM LHYDPNKRIS AKAALAHPFF QDVTKPVPHL RL

+
Macromolecule #17: Cyclin A2

MacromoleculeName: Cyclin A2 / type: protein_or_peptide / ID: 17 / Enantiomer: LEVO
SequenceString: MLGNSAPGPA TREAGSALLA LQQTALQEDQ ENINPEKAAP VQQPRTRAAL AVLKSGNPRG LAQQQRPKT RRVAPLKDLP VNDEHVTVPP WKANSKQPAF TIHVDEAEKE AQKKPAESQK I EREDALAF NSAISLPGPR KPLVPLDYPM DGSFESPHTM DMSIILEDEK ...String:
MLGNSAPGPA TREAGSALLA LQQTALQEDQ ENINPEKAAP VQQPRTRAAL AVLKSGNPRG LAQQQRPKT RRVAPLKDLP VNDEHVTVPP WKANSKQPAF TIHVDEAEKE AQKKPAESQK I EREDALAF NSAISLPGPR KPLVPLDYPM DGSFESPHTM DMSIILEDEK PVSVNEVPDY HE DIHTYLR EMEVKCKPKV GYMKKQPDIT NSMRAILVDW LVEVGEEYKL QNETLHLAVN YID RFLSSM SVLRGKLQLV GTAAMLLASK FEEIYPPEVA EFVYITDDTY TKKQVLRMEH LVLK VLTFD LAAPTVNQFL TQYFLHQQPA NCKVESLAMF LGELSLIDAD PYLKYLPSVI AGAAF HLAL YTVTGQSWPE SLIRKTGYTL ESLKPCLMDL HQTYLKAPQH AQQSIREKYK NSKYHG VSL LNPPETLNL

+
Macromolecule #18: Cks2

MacromoleculeName: Cks2 / type: protein_or_peptide / ID: 18 / Enantiomer: LEVO
SequenceString:
MAHKQIYYSD KYFDEHYEYR HVMLPRELSK QVPKTHLMSE EEWRRLGVQQ SLGWVHYMIH EPEPHILLF RRPLPKDQQK

+
Macromolecule #19: BubR1

MacromoleculeName: BubR1 / type: protein_or_peptide / ID: 19 / Enantiomer: LEVO
SequenceString: MAAVKKEGGA LSEAMSLEGD EWELSKENVQ PLRQGRIMST LQGALAQESA CNNTLQQQKR AFEYEIRFY TGNDPLDVWD RYISWTEQNY PQGGKESNMS TLLERAVEAL QGEKRYYSDP R FLNLWLKL GRLCNEPLDM YSYLHNQGIG VSLAQFYISW AEEYEARENF ...String:
MAAVKKEGGA LSEAMSLEGD EWELSKENVQ PLRQGRIMST LQGALAQESA CNNTLQQQKR AFEYEIRFY TGNDPLDVWD RYISWTEQNY PQGGKESNMS TLLERAVEAL QGEKRYYSDP R FLNLWLKL GRLCNEPLDM YSYLHNQGIG VSLAQFYISW AEEYEARENF RKADAIFQEG IQ QKAEPLE RLQSQHRQFQ ARVSRQTLLA LEKEEEEEVF ESSVPQRSTL AELKSKGKKT ARA PIIRVG GALKAPSQNR GLQNPFPQQM QNNSRITVFD ENADEASTAE LSKPTVQPWI APPM PRAKE NELQAGPWNT GRSLEHRPRG NTASLIAVPA VLPSFTPYVE ETARQPVMTP CKIEP SINH ILSTRKPGKE EGDPLQRVQS HQQASEEKKE KMMYCKEKIY AGVGEFSFEE IRAEVF RKK LKEQREAELL TSAEKRAEMQ KQIEEMEKKL KEIQTTQQER TGDQQEETMP TKETTKL QI ASESQKIPGM TLSSSVCQVN CCARETSLAE NIWQEQPHSK GPSVPFSIFD EFLLSEKK N KSPPADPPRV LAQRRPLAVL KTSESITSNE DVSPDVCDEF TGIEPLSEDA IITGFRNVT ICPNPEDTCD FARAARFVST PFHEIMSLKD LPSDPERLLP EEDLDVKTSE DQQTACGTIY SQTLSIKKL SPIIEDSREA THSSGFSGSS ASVASTSSIK CLQIPEKLEL TNETSENPTQ S PWCSQYRR QLLKSLPELS ASAELCIEDR PMPKLEIEKE IELGNEDYCI KREYLICEDY KL FWVAPRN SAELTVIKVS SQPVPWDFYI NLKLKERLNE DFDHFCSCYQ YQDGCIVWHQ YIN CFTLQD LLQHSEYITH EITVLIIYNL LTIVEMLHKA EIVHGDLSPR CLILRNRIHD PYDC NKNNQ ALKIVDFSYS VDLRVQLDVF TLSGFRTVQI LEGQKILANC SSPYQVDLFG IADLA HLLL FKEHLQVFWD GSFWKLSQNI SELKDGELWN KFFVRILNAN DEATVSVLGE LAAEMN GVF DTTFQSHLNK ALWKVGKLTS PGALLFQ

+
Macromolecule #20: Mad2

MacromoleculeName: Mad2 / type: protein_or_peptide / ID: 20 / Enantiomer: LEVO
SequenceString: MALQLSREQG ITLRGSAEIV AEFFSFGINS ILYQRGIYPS ETFTRVQKYG LTLLVTTDLE LIKYLNNVV EQLKDWLYKC SVQKLVVVIS NIESGEVLER WQFDIECDKT AKDDSAPREK S QKAIQDEI RSVIRQITAT VTFLPLLEVS CSFDLLIYTD KDLVVPEKWE ...String:
MALQLSREQG ITLRGSAEIV AEFFSFGINS ILYQRGIYPS ETFTRVQKYG LTLLVTTDLE LIKYLNNVV EQLKDWLYKC SVQKLVVVIS NIESGEVLER WQFDIECDKT AKDDSAPREK S QKAIQDEI RSVIRQITAT VTFLPLLEVS CSFDLLIYTD KDLVVPEKWE ESGPQFITNS EE VRLRSFT TTIHKVNSMV AYKIPVND

+
Macromolecule #21: Bub3

MacromoleculeName: Bub3 / type: protein_or_peptide / ID: 21 / Enantiomer: LEVO
SequenceString: MTGSNEFKLN QPPEDGISSV KFSPNTSQFL LVSSWDTSVR LYDVPANSMR LKYQHTGAVL DCAFYDPTH AWSGGLDHQL KMHDLNTDQE NLVGTHDAPI RCVEYCPEVN VMVTGSWDQT V KLWDPRTP CNAGTFSQPE KVYTLSVSGD RLIVGTAGRR VLVWDLRNMG ...String:
MTGSNEFKLN QPPEDGISSV KFSPNTSQFL LVSSWDTSVR LYDVPANSMR LKYQHTGAVL DCAFYDPTH AWSGGLDHQL KMHDLNTDQE NLVGTHDAPI RCVEYCPEVN VMVTGSWDQT V KLWDPRTP CNAGTFSQPE KVYTLSVSGD RLIVGTAGRR VLVWDLRNMG YVQQRRESSL KY QTRCIRA FPNKQGYVLS SIEGRVAVEY LDPSPEVQKK KYAFKCHRLK ENNIEQIYPV NAI SFHNIH NTFATGGSDG FVNIWDPFNK KRLCQFHRYP TSIASLAFSN DGTTLAIASS YMYE MDDTE HPEDGIFIRQ VTDAETKPKS PCT

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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

Concentration0.15 mg/mL
BufferpH: 8 / Details: 20mM Hepes, 150mM NaCl, 0.5mM TCEP
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeFEI POLARA 300
Image recordingFilm or detector model: FEI FALCON III (4k x 4k) / Detector mode: INTEGRATING / Average electron dose: 28.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD
Experimental equipment
Model: Tecnai Polara / Image courtesy: FEI Company

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Image processing

Final reconstructionApplied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 3.7 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 3.0) / Number images used: 81207
Initial angle assignmentType: MAXIMUM LIKELIHOOD
Final angle assignmentType: MAXIMUM LIKELIHOOD
FSC plot (resolution estimation)

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