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- EMDB-4467: Cryo-EM structure of the APC/C-Cdc20-Cdk2-cyclinA2delD1-Cks2 comp... -

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Basic information

Entry
Database: EMDB / ID: EMD-4467
TitleCryo-EM structure of the APC/C-Cdc20-Cdk2-cyclinA2delD1-Cks2 complex (with the D1 box mutated)
Map dataunsharpened map
Sample
  • Complex: APC/C-Cdc20-Cdk2-cyclinA2delD1-Cks2 complex (with the D1 box mutated)
    • Protein or peptide: x 18 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: 0.014
  • Imaged by UCSF Chimera
  • Download
  • Surface view colored by radius
  • Surface level: 0.014
  • Imaged by UCSF Chimera
  • Download
Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

emd_4467.png

Downloads & links

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Map

FileDownload / File: emd_4467.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Annotationunsharpened map
Voxel sizeX=Y=Z: 1.1 Å
Density
Contour LevelBy AUTHOR: 0.014 / Movie #1: 0.014
Minimum - Maximum-0.01246849 - 0.044519883
Average (Standard dev.)0.00003197904 (±0.0019779908)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions400400400
Spacing400400400
CellA=B=C: 440.0 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z1.11.11.1
M x/y/z400400400
origin x/y/z0.0000.0000.000
length x/y/z440.000440.000440.000
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-0.0120.0450.000

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Supplemental data

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Additional map: sharpened map with a B factor -50

Fileemd_4467_additional.map
Annotationsharpened map with a B factor -50
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Additional map: sharpened map with a B factor -50

Fileemd_4467_additional_1.map
Annotationsharpened map with a B factor -50
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: halfmap1

Fileemd_4467_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_4467_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 : APC/C-Cdc20-Cdk2-cyclinA2delD1-Cks2 complex (with the D1 box mutated)

EntireName: APC/C-Cdc20-Cdk2-cyclinA2delD1-Cks2 complex (with the D1 box mutated)
Components
  • Complex: APC/C-Cdc20-Cdk2-cyclinA2delD1-Cks2 complex (with the D1 box mutated)
    • 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: Cdk2Cyclin-dependent kinase 2
    • Protein or peptide: Cyclin A2
    • Protein or peptide: Cks2

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Supramolecule #1: APC/C-Cdc20-Cdk2-cyclinA2delD1-Cks2 complex (with the D1 box mutated)

SupramoleculeName: APC/C-Cdc20-Cdk2-cyclinA2delD1-Cks2 complex (with the D1 box mutated)
type: complex / ID: 1 / Parent: 0 / Macromolecule list: all
Source (natural)Organism: Homo sapiens (human)
Recombinant expressionOrganism: unidentified baculovirus
Molecular weightTheoretical: 1.3 MDa

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Macromolecule #1: Apc1

MacromoleculeName: Apc1 / type: protein_or_peptide / ID: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Recombinant expressionOrganism: unidentified baculovirus
SequenceString: MSNFYEERTT MIAARDLQEF VPFGRDHCKH HPNALNLQLR QLQPASELWS SDGAAGLVGS LQEVTIHEK QKESWQLRKG VSEIGEDVDY DEELYVAGNM VIWSKGSKSQ ALAVYKAFTV D SPVQQALW CDFIISQDKS EKAYSSNEVE KCICILQSSC INMHSIEGKD ...String:
MSNFYEERTT MIAARDLQEF VPFGRDHCKH HPNALNLQLR QLQPASELWS SDGAAGLVGS LQEVTIHEK QKESWQLRKG VSEIGEDVDY DEELYVAGNM VIWSKGSKSQ ALAVYKAFTV D SPVQQALW CDFIISQDKS EKAYSSNEVE KCICILQSSC INMHSIEGKD YIASLPFQVA NV WPTKYGL LFERSASSHE VPPGSPREPL PTMFSMLHPL DEITPLVCKS GSLFGSSRVQ YVV DHAMKI VFLNTDPSIV MTYDAVQNVH SVWTLRRVKS EEENVVLKFS EQGGTPQNVA TSSS LTAHL RLAPETEPIV PELCIDHLWT ETITNI REK NSQASKVFIT SDLCGQKFLC FLVESQLQLR CVKFQESNDK TQLIFGSVTN IPAKDAA PV EKIDTMLVLE GSGNLVLYTG VVRVGKVFIP GLPAPSLTMS NTMPRPSTPL DGVSTPKP L SKLLGSLDEV VLLSPVPELR DSSKLHDSLY NEDCTFQQLG TYIHSIRDPV HNRVTLELS NGSMVRITIP EIATSELVQT CLQAIKFILP KEIAVQMLVK WYNVHSAPGG PSYHSEWNLF VTCLMNMMG YNTDRLAWTR NFDFEGSLSP VIAPKKARPS ETGSDDDWEY LLNSDYHQNV E SHLLNRSL CLSPSEASQM KDEDFSQNLS LDSSTLLFTH IPAIFFVLHL VYEELKLNTL MG EGICSLV ELLVQLARDL KLGPYVDHYY RDYPTLVRTT GQVCTIDPGQ TGFMHHPSFF TSE PPSIYQ WVSSCLKGEG MPPYPYLPGI CERSRLVVLS IALYILGDES LVSDESSQYL TRIT IAPQK LQVEQEENRF SFRHSTSVSS LAERLVVWMT NVGFTLRDLE TLPFGIALPI RDAIY HCRE QPASDWPEAV CLLIGRQDLS KQACEGNLPK GKSVLSSDVP SGTETEEEDD GMNDMN HEV MSLIWSEDLR VQDVRRLLQS AHPVRVNVVQ YPELSDHEFI EEKENRLLQL CQRTMAL PV GRGMFTLFSY HPVPTEPLPI PKLNLTGRAP PRNTTVDLNS GNIDVPPNMT SWASFHNG V AAGLKIAPAS QIDSAWIVYN KPKHAELANE YAGFLMALGL NGHLTKLATL NIHDYLTKG HEMTSIGLLL GVSAAKLGTM DMSITRLLSI HIPALLPPTS TELDVPHNVQ VAAVVGIGLV YQGTAHRHT AEVLLAEIGR PPGPEMEYCT DRESYSLAAG LALGMVCLGH GSNLIGMSDL N VPEQLYQY MVGGHRRFQT GMHREKHKSP SYQIKEGDTI NVDVTCPGAT LALAMIYLKT NN RSIADWL RAPDTMYLLD FVKPEFLLLR TLARCLILWD DILPNSKWVD SNVPQIIREN SIS LSEIEL PCSEDLNLET LSQAHVYIIA GACLSLGFRF AGSENLSAFN CLHKFAKDFM TYLS APNAS VTGPHNLETC LSVVLLSLAM VMAGSGNLKV LQLCRFLHMK TGGEMNYGFH LAHHM ALGL LFLGGGRYSL STSNSSIAAL LCALYPHFPA HSTDNRYHLQ ALRHLYVLAA EPRLLV PVD VDTNTPCYAL LEVTYKGTQW YEQTKEELMA PTLLPELHLL KQIKVKGPRY WELLIDL SK GTQHLKSILS KDGVLYVKLR AGQLSYKEDP MGWQSLLAQT VANRNSEARA FKPETISA F TSDPALLSFA EYFCKPTVNM GQKQEILDLF SSVLYECVTQ ETPEMLPAYI AMDQAIRRL GRREMSETSE LWQIKLVLEF FSSRSHQERL QNHPKRGLFM NSEFLPVVKC TIDNTLDQWL QVGGDMCVH AYLSGQPLEE SQLSMLACFL VYHSVPAPQH LPPIGLEGST SFAELLFKFK Q LKMPVRAL LRLAPLLLGN PQPMVM

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Macromolecule #2: Apc2

MacromoleculeName: Apc2 / type: protein_or_peptide / ID: 2 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Recombinant expressionOrganism: unidentified baculovirus
SequenceString: MAAAVVVAEG DSDSRPGQEL LVAWNTVSTG LVPPAALGLV SSRTSGAVPP KEEELRAAVE VLRGHGLHS VLEEWFVEVL QNDLQANISP EFWNAISQCE NSADEPQCLL LLLDAFGLLE S RLDPYLRS LELLEKWTRL GLLMGTGAQG LREEVHTMLR GVLFFSTPRT ...String:
MAAAVVVAEG DSDSRPGQEL LVAWNTVSTG LVPPAALGLV SSRTSGAVPP KEEELRAAVE VLRGHGLHS VLEEWFVEVL QNDLQANISP EFWNAISQCE NSADEPQCLL LLLDAFGLLE S RLDPYLRS LELLEKWTRL GLLMGTGAQG LREEVHTMLR GVLFFSTPRT FQEMIQRLYG CF LRVYMQS KRKGEGGTDP ELEGELDSRY ARRRYYRLLQ SPLCAGCSSD KQQCWCRQAL EQF HQLSQV LHRLSLLERV SAEAVTTTLH QVTRERMEDR CRGEYERSFL REFHKWIERV VGWL GKVFL QDGPARPASP EAGNTLRRWR CHVQRFFYRI YASLRIEELF SIVRDFPDSR PAIED LKYC LERTDQRQQL LVSLKAALET RLLHPGVNTC DIITLYISAI KALRVLDPSM VILEVA CEP IRRYLRTRED TVRQIVAGLT GDSDGTGDLA VELSKTDPAS LETGQDSEDD SGEPEDW VP DPVDADPGKS SSKRRSSDII SLLVSIYGSK DLFINEYRSL LADRLLHQFS FSPEREIR N VELLKLRFGE APMHFCEVML KDMADSRRIN ANIREEDEKR PAEEQPPFGV YAVILSSEF WPPFKDEKLE VPEDIRAALE AYCKKYEQLK AMRTLSWKHT LGLVTMDVEL ADRTLSVAVT PVQAVILLY FQDQASWTLE ELSKAVKMPV ALLRRRMSVW LQQGVLREEP PGTFSVIEEE R PQDRDNMV LIDSDDESDS GMASQADQKE EELLLFWTYI QAMLTNLESL SLDRIYNMLR MF VVTGPAL AEIDLQELQG YLQKKVRDQQ LVYSAGVYRL PKNCS

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Macromolecule #3: Apc3

MacromoleculeName: Apc3 / type: protein_or_peptide / ID: 3 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Recombinant expressionOrganism: unidentified baculovirus
SequenceString: MTVLQEPVQA AIWQALNHYA YRDAVFLAER LYAEVHSEEA LFLLATCYYR SGKAYKAYRL LKGHSCTTP QCKYLLAKCC VDLSKLAEGE QILSGGVFNK QKSHDDIVTE FGDSACFTLS L LGHVYCKT DRLAKGSECY QKSLSLNPFL WSPFESLCEI GEKPDPDQTF ...String:
MTVLQEPVQA AIWQALNHYA YRDAVFLAER LYAEVHSEEA LFLLATCYYR SGKAYKAYRL LKGHSCTTP QCKYLLAKCC VDLSKLAEGE QILSGGVFNK QKSHDDIVTE FGDSACFTLS L LGHVYCKT DRLAKGSECY QKSLSLNPFL WSPFESLCEI GEKPDPDQTF KFTSLQNFSN CL PNSCTTQ VPNHSLSHRQ PETVLTETPQ DTIELNRLNL ESSNSKYSLN TDSSVSYIDS AVI SPDTVP LGTGTSILSK QVQNKPKTGR SLLGGPAALS PLTPSFGILP LETPSPGDGS YLQN YTNTP PVIDVPSTGA PSKKSVARIG QTGTKSVFSQ SGNSREVTPI LAQTQSSGPQ TSTTP QVLS PTITSPPNAL PRRSSRLFTS DSSTTKENSK KLKMKFPPKI PNRKTKSKTN KGGITQ PNI NDSLEITKLD SSIISEGKIS TITPQIQAFN LQKAAAEGLM SLLREMGKGY LALCSYN CK EAINILSHLP SHHYNTGWVL CQIGRAYFEL SEYMQAERIF SEVRRIENYR VEGMEIYS T TLWHLQKDVA LSVLSKDLTD MDKNSPEAWC AAGNCFSLQR EHDIAIKFFQ RAIQVDPNY AYAYTLLGHE FVLTEELDKA LACFRNAIRV NPRHYNAWYG LGMIYYKQEK FSLAEMHFQK ALDINPQSS VLLCHIGVVQ HALKKSEKAL DTLNKAIVID PKNPLCKFHR ASVLFANEKY K SALQELEE LKQIVPKESL VYFLIGKVYK KLGQTHLALM NFSWAMDLDP KGANNQIKEA ID KRYLPDD EEPITQEEQI MGTDESQESS MTDADDTQLH AAESDEF

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Macromolecule #4: Apc4

MacromoleculeName: Apc4 / type: protein_or_peptide / ID: 4 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Recombinant expressionOrganism: unidentified baculovirus
SequenceString: MLRFPTCFPS FRVVGEKQLP QEIIFLVWSP KRDLIALANT AGEVLLHRLA SFHRVWSFPP NENTGKEVT CLAWRPDGKL LAFALADTKK IVLCDVEKPE SLHSFSVEAP VSCMHWMEVT V ESSVLTSF YNAEDESNLL LPKLPTLPKN YSNTSKIFSE ENSDEIIKLL ...String:
MLRFPTCFPS FRVVGEKQLP QEIIFLVWSP KRDLIALANT AGEVLLHRLA SFHRVWSFPP NENTGKEVT CLAWRPDGKL LAFALADTKK IVLCDVEKPE SLHSFSVEAP VSCMHWMEVT V ESSVLTSF YNAEDESNLL LPKLPTLPKN YSNTSKIFSE ENSDEIIKLL GDVRLNILVL GG SSGFIEL YAYGMFKIAR VTGIAGTCLA LCLSSDLKSL SVVTEVSTNG ASEVSYFQLE TNL LYSFLP EVTRMARKFT HISALLQYIN LSLTCMCEAW EEILMQMDSR LTKFVQEKNT TTSV QDEFM HLLLWGKASA ELQTLLMNQL TVKGLKKLGQ SIESSYSSIQ KLVISHLQSG SESLL YHLS ELKGMASWKQ KYEPLGLDAA GIEEAITAVG SFILKANELL QVIDSSMKNF KAFFRW LYV AMLRMTEDHV LPELNKMTQK DITFVAEFLT EHFNEAPDLY NRKGKYFNVE RVGQYLK DE DDDLVSPPNT EGNQWYDFLQ NSSHLKESPL LFPYYPRKSL HFVKRRMENI IDQCLQKP A DVIGKSMNQA ICIPLYRDTR SEDSTRRLFK FPFLWNNKTS NLHYLLFTIL EDSLYKMCI LRRHTDISQS VSNGLIAIKF GSFTYATTEK VRRSIYSCLD AQFYDDETVT VVLKDTVGRE GRDRLLVQL PLSLVYNSED SAEYQFTGTY STRLDEQCSA IPTRTMHFEK HWRLLESMKA Q YVAGNGFR KVSCVLSSNL RHVRVFEMDI DDEWELDESS DEEEEASNKP VKIKEEVLSE SE AENQQAG AAALAPEIVI KVEKLDPELD S

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Macromolecule #5: Apc5

MacromoleculeName: Apc5 / type: protein_or_peptide / ID: 5 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Recombinant expressionOrganism: unidentified baculovirus
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
Source (natural)Organism: Homo sapiens (human)
Recombinant expressionOrganism: unidentified baculovirus
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
Source (natural)Organism: Homo sapiens (human)
Recombinant expressionOrganism: unidentified baculovirus
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
Source (natural)Organism: Homo sapiens (human)
Recombinant expressionOrganism: unidentified baculovirus
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
Source (natural)Organism: Homo sapiens (human)
Recombinant expressionOrganism: unidentified baculovirus
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
Source (natural)Organism: Homo sapiens (human)
Recombinant expressionOrganism: unidentified baculovirus
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
Source (natural)Organism: Homo sapiens (human)
Recombinant expressionOrganism: unidentified baculovirus
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
Source (natural)Organism: Homo sapiens (human)
Recombinant expressionOrganism: unidentified baculovirus
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
Source (natural)Organism: Homo sapiens (human)
Recombinant expressionOrganism: unidentified baculovirus
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
Source (natural)Organism: Homo sapiens (human)
Recombinant expressionOrganism: unidentified baculovirus
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
Source (natural)Organism: Homo sapiens (human)
Recombinant expressionOrganism: unidentified baculovirus
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

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Macromolecule #16: Cdk2

MacromoleculeName: Cdk2 / type: protein_or_peptide / ID: 16 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Recombinant expressionOrganism: Escherichia coli (E. coli)
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

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Macromolecule #17: Cyclin A2

MacromoleculeName: Cyclin A2 / type: protein_or_peptide / ID: 17 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Recombinant expressionOrganism: Escherichia coli (E. coli)
SequenceString: MLGNSAPGPA TREAGSALLA LQQTALQEDQ ENINPEKAAP VQQPRTAAAA AVAKSGNPRG LAQQQRPKT RRVAPLKDLP VNDEHVTVPP WKANSKQPAF TIHVDEAEKE AQKKPAESQK I EREDALAF NSAISLPGPR KPLVPLDYPM DGSFESPHTM DMSIILEDEK ...String:
MLGNSAPGPA TREAGSALLA LQQTALQEDQ ENINPEKAAP VQQPRTAAAA AVAKSGNPRG 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

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Macromolecule #18: Cks2

MacromoleculeName: Cks2 / type: protein_or_peptide / ID: 18 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Recombinant expressionOrganism: Escherichia coli (E. coli)
SequenceString:
MAHKQIYYSD KYFDEHYEYR HVMLPRELSK QVPKTHLMSE EEWRRLGVQQ SLGWVHYMIH EPEPHILLF RRPLPKDQQK

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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
GridModel: Quantifoil R3.5/1 / Material: COPPER / Support film - Material: CARBON
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
Image recordingFilm or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 28.0 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 reconstructionApplied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 3.7 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 61415
FSC plot (resolution estimation)

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