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Yorodumi- EMDB-15284: Cryo-EM structure of USP1-UAF1 bound to FANCI and mono-ubiquitina... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-15284 | |||||||||
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Title | Cryo-EM structure of USP1-UAF1 bound to FANCI and mono-ubiquitinated FANCD2 without ML323 (consensus reconstruction) | |||||||||
Map data | Globally sharpened map | |||||||||
Sample |
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Function / homology | Function and homology information positive regulation of error-prone translesion synthesis / regulation of protein monoubiquitination / Signaling by cytosolic PDGFRA and PDGFRB fusion proteins / regulation of CD40 signaling pathway / regulation of regulatory T cell differentiation / monoubiquitinated protein deubiquitination / double-strand break repair involved in meiotic recombination / homologous chromosome pairing at meiosis / gamete generation / neuronal stem cell population maintenance ...positive regulation of error-prone translesion synthesis / regulation of protein monoubiquitination / Signaling by cytosolic PDGFRA and PDGFRB fusion proteins / regulation of CD40 signaling pathway / regulation of regulatory T cell differentiation / monoubiquitinated protein deubiquitination / double-strand break repair involved in meiotic recombination / homologous chromosome pairing at meiosis / gamete generation / neuronal stem cell population maintenance / brain morphogenesis / deubiquitinase activator activity / DNA repair complex / skeletal system morphogenesis / mitotic intra-S DNA damage checkpoint signaling / skin development / seminiferous tubule development / protein deubiquitination / Regulation of pyruvate metabolism / Regulation of TBK1, IKKε (IKBKE)-mediated activation of IRF3, IRF7 / Regulation of TBK1, IKKε-mediated activation of IRF3, IRF7 upon TLR3 ligation / positive regulation of double-strand break repair via homologous recombination / homeostasis of number of cells / single fertilization / embryonic organ development / regulation of DNA repair / interstrand cross-link repair / DNA polymerase binding / response to UV / condensed chromosome / Maturation of protein E / Maturation of protein E / ER Quality Control Compartment (ERQC) / Myoclonic epilepsy of Lafora / FLT3 signaling by CBL mutants / Prevention of phagosomal-lysosomal fusion / IRAK2 mediated activation of TAK1 complex / Alpha-protein kinase 1 signaling pathway / Glycogen synthesis / IRAK1 recruits IKK complex / IRAK1 recruits IKK complex upon TLR7/8 or 9 stimulation / Membrane binding and targetting of GAG proteins / Endosomal Sorting Complex Required For Transport (ESCRT) / IRAK2 mediated activation of TAK1 complex upon TLR7/8 or 9 stimulation / PTK6 Regulates RTKs and Their Effectors AKT1 and DOK1 / Negative regulation of FLT3 / Constitutive Signaling by NOTCH1 HD Domain Mutants / Regulation of FZD by ubiquitination / TICAM1,TRAF6-dependent induction of TAK1 complex / NOTCH2 Activation and Transmission of Signal to the Nucleus / TICAM1-dependent activation of IRF3/IRF7 / APC/C:Cdc20 mediated degradation of Cyclin B / p75NTR recruits signalling complexes / Downregulation of ERBB4 signaling / APC-Cdc20 mediated degradation of Nek2A / PINK1-PRKN Mediated Mitophagy / TRAF6-mediated induction of TAK1 complex within TLR4 complex / TRAF6 mediated IRF7 activation in TLR7/8 or 9 signaling / Pexophagy / Regulation of innate immune responses to cytosolic DNA / InlA-mediated entry of Listeria monocytogenes into host cells / VLDLR internalisation and degradation / Downregulation of ERBB2:ERBB3 signaling / NF-kB is activated and signals survival / NRIF signals cell death from the nucleus / Regulation of PTEN localization / Activated NOTCH1 Transmits Signal to the Nucleus / Regulation of BACH1 activity / Translesion synthesis by REV1 / Synthesis of active ubiquitin: roles of E1 and E2 enzymes / Translesion synthesis by POLK / MAP3K8 (TPL2)-dependent MAPK1/3 activation / TICAM1, RIP1-mediated IKK complex recruitment / Downregulation of TGF-beta receptor signaling / Activation of IRF3, IRF7 mediated by TBK1, IKKε (IKBKE) / Translesion synthesis by POLI / Gap-filling DNA repair synthesis and ligation in GG-NER / Josephin domain DUBs / Regulation of activated PAK-2p34 by proteasome mediated degradation / InlB-mediated entry of Listeria monocytogenes into host cell / IKK complex recruitment mediated by RIP1 / JNK (c-Jun kinases) phosphorylation and activation mediated by activated human TAK1 / TGF-beta receptor signaling in EMT (epithelial to mesenchymal transition) / N-glycan trimming in the ER and Calnexin/Calreticulin cycle / Autodegradation of Cdh1 by Cdh1:APC/C / TNFR1-induced NF-kappa-B signaling pathway / APC/C:Cdc20 mediated degradation of Securin / ubiquitin binding / positive regulation of protein ubiquitination / Asymmetric localization of PCP proteins / TCF dependent signaling in response to WNT / SCF-beta-TrCP mediated degradation of Emi1 / NIK-->noncanonical NF-kB signaling / skeletal system development / Ubiquitin-dependent degradation of Cyclin D / Regulation of NF-kappa B signaling / positive regulation of epithelial cell proliferation / AUF1 (hnRNP D0) binds and destabilizes mRNA / TNFR2 non-canonical NF-kB pathway / activated TAK1 mediates p38 MAPK activation Similarity search - Function | |||||||||
Biological species | Homo sapiens (human) / synthetic construct (others) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 2.8 Å | |||||||||
Authors | Rennie ML / Walden H | |||||||||
Funding support | European Union, United Kingdom, 2 items
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Citation | Journal: Sci Adv / Year: 2022 Title: Cryo-EM reveals a mechanism of USP1 inhibition through a cryptic binding site. Authors: Martin L Rennie / Connor Arkinson / Viduth K Chaugule / Helen Walden / Abstract: Repair of DNA damage is critical to genomic integrity and frequently disrupted in cancers. Ubiquitin-specific protease 1 (USP1), a nucleus-localized deubiquitinase, lies at the interface of multiple ...Repair of DNA damage is critical to genomic integrity and frequently disrupted in cancers. Ubiquitin-specific protease 1 (USP1), a nucleus-localized deubiquitinase, lies at the interface of multiple DNA repair pathways and is a promising drug target for certain cancers. Although multiple inhibitors of this enzyme, including one in phase 1 clinical trials, have been established, their binding mode is unknown. Here, we use cryo-electron microscopy to study an assembled enzyme-substrate-inhibitor complex of USP1 and the well-established inhibitor, ML323. Achieving 2.5-Å resolution, with and without ML323, we find an unusual binding mode in which the inhibitor disrupts part of the hydrophobic core of USP1. The consequent conformational changes in the secondary structure lead to subtle rearrangements in the active site that underlie the mechanism of inhibition. These structures provide a platform for structure-based drug design targeting USP1. | |||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_15284.map.gz | 118.1 MB | EMDB map data format | |
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Header (meta data) | emd-15284-v30.xml emd-15284.xml | 28 KB 28 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_15284_fsc.xml | 11.9 KB | Display | FSC data file |
Images | emd_15284.png | 181.5 KB | ||
Masks | emd_15284_msk_1.map | 125 MB | Mask map | |
Others | emd_15284_half_map_1.map.gz emd_15284_half_map_2.map.gz | 116 MB 116 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-15284 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-15284 | HTTPS FTP |
-Validation report
Summary document | emd_15284_validation.pdf.gz | 821.5 KB | Display | EMDB validaton report |
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Full document | emd_15284_full_validation.pdf.gz | 821.1 KB | Display | |
Data in XML | emd_15284_validation.xml.gz | 19 KB | Display | |
Data in CIF | emd_15284_validation.cif.gz | 24.3 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-15284 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-15284 | HTTPS FTP |
-Related structure data
Related structure data | 8a9jMC 7zh3C 7zh4C 8a9kC C: citing same article (ref.) M: atomic model generated by this map |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_15284.map.gz / Format: CCP4 / Size: 125 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||
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Annotation | Globally sharpened map | ||||||||||||||||||||
Voxel size | X=Y=Z: 1.06 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Mask #1
File | emd_15284_msk_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #1
File | emd_15284_half_map_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #2
File | emd_15284_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Sample components
-Entire : USP1(C90S)-UAF1 bound to FANCI and mono-ubiquitinated FANCD2 with...
Entire | Name: USP1(C90S)-UAF1 bound to FANCI and mono-ubiquitinated FANCD2 with dsDNA |
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Components |
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-Supramolecule #1: USP1(C90S)-UAF1 bound to FANCI and mono-ubiquitinated FANCD2 with...
Supramolecule | Name: USP1(C90S)-UAF1 bound to FANCI and mono-ubiquitinated FANCD2 with dsDNA type: complex / Chimera: Yes / ID: 1 / Parent: 0 / Macromolecule list: #1-#6 |
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-Supramolecule #2: Fanconi anemia group I protein, Fanconi anemia group D2 protein w...
Supramolecule | Name: Fanconi anemia group I protein, Fanconi anemia group D2 protein with ubiquitin conjugated to K561, Ubiquitin carboxyl-terminal hydrolase 1, WD repeat-containing protein 48 type: complex / Chimera: Yes / ID: 2 / Parent: 1 / Macromolecule list: #1-#6 |
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Source (natural) | Organism: Homo sapiens (human) |
Recombinant expression | Organism: Spodoptera frugiperda (fall armyworm) |
-Macromolecule #1: Fanconi anemia group I protein
Macromolecule | Name: Fanconi anemia group I protein / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 150.459125 KDa |
Recombinant expression | Organism: Spodoptera frugiperda (fall armyworm) |
Sequence | String: MHHHHHHMDQ KILSLAAEKT ADKLQEFLQT LREGDLTNLL QNQAVKGKVA GALLRAIFKG SPCSEEAGTL RRRKIYTCCI QLVESGDLQ KEIASEIIGL LMLEAHHFPG PLLVELANEF ISAVREGSLV NGKSLELLPI ILTALATKKE NLAYGKGVLS G EECKKQLI ...String: MHHHHHHMDQ KILSLAAEKT ADKLQEFLQT LREGDLTNLL QNQAVKGKVA GALLRAIFKG SPCSEEAGTL RRRKIYTCCI QLVESGDLQ KEIASEIIGL LMLEAHHFPG PLLVELANEF ISAVREGSLV NGKSLELLPI ILTALATKKE NLAYGKGVLS G EECKKQLI NTLCSGRWDQ QYVIQLTSMF KDVPLTAEEV EFVVEKALSM FSKMNLQEIP PLVYQLLVLS SKGSRKSVLE GI IAFFSAL DKQHNEEQSG DELLDVVTVP SGELRHVEGT IILHIVFAIK LDYELGRELV KHLKVGQQGD SNNNLSPFSI ALL LSVTRI QRFQDQVLDL LKTSVVKSFK DLQLLQGSKF LQNLVPHRSY VSTMILEVVK NSVHSWDHVT QGLVELGFIL MDSY GPKKV LDGKTIETSP SLSRMPNQHA CKLGANILLE TFKIHEMIRQ EILEQVLNRV VTRASSPISH FLDLLSNIVM YAPLV LQSC SSKVTEAFDY LSFLPLQTVQ RLLKAVQPLL KVSMSMRDCL ILVLRKAMFA NQLDARKSAV AGFLLLLKNF KVLGSL SSS QCSQSLSVSQ VHVDVHSHYN SVANETFCLE IMDSLRRCLS QQADVRLMLY EGFYDVLRRN SQLANSVMQT LLSQLKQ FY EPKPDLLPPL KLEACILTQG DKISLQEPLD YLLCCIQHCL AWYKNTVIPL QQGEEEEEEE EAFYEDLDDI LESITNRM I KSELEDFELD KSADFSQSTS IGIKNNICAF LVMGVCEVLI EYNFSISSFS KNRFEDILSL FMCYKKLSDI LNEKAGKAK TKMANKTSDS LLSMKFVSSL LTALFRDSIQ SHQESLSVLR SSNEFMRYAV NVALQKVQQL KETGHVSGPD GQNPEKIFQN LCDITRVLL WRYTSIPTSV EESGKKEKGK SISLLCLEGL QKIFSAVQQF YQPKIQQFLR ALDVTDKEGE EREDADVSVT Q RTAFQIRQ FQRSLLNLLS SQEEDFNSKE ALLLVTVLTS LSKLLEPSSP QFVQMLSWTS KICKENSRED ALFCKSLMNL LF SLHVSYK SPVILLRDLS QDIHGHLGDI DQDVEVEKTN HFAIVNLRTA APTVCLLVLS QAEKVLEEVD WLITKLKGQV SQE TLSEEA SSQATLPNQP VEKAIIMQLG TLLTFFHELV QTALPSGSCV DTLLKDLCKM YTTLTALVRY YLQVCQSSGG IPKN MEKLV KLSGSHLTPL CYSFISYVQN KSKSLNYTGE KKEKPAAVAT AMARVLRETK PIPNLIFAIE QYEKFLIHLS KKSKV NLMQ HMKLSTSRDF KIKGNILDMV LREDGEDENE EGTASEHGGQ NKEPAKKKRK K |
-Macromolecule #2: Fanconi anemia group D2 protein
Macromolecule | Name: Fanconi anemia group D2 protein / type: protein_or_peptide / ID: 2 / Details: Ubiquitin conjugated to K561 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 164.623828 KDa |
Recombinant expression | Organism: Spodoptera frugiperda (fall armyworm) |
Sequence | String: GPGSMVSKRR LSKSEDKESL TEDASKTRKQ PLSKKTKKSH IANEVEENDS IFVKLLKISG IILKTGESQN QLAVDQIAFQ KKLFQTLRR HPSYPKIIEE FVSGLESYIE DEDSFRNCLL SCERLQDEEA SMGASYSKSL IKLLLGIDIL QPAIIKTLFE K LPEYFFEN ...String: GPGSMVSKRR LSKSEDKESL TEDASKTRKQ PLSKKTKKSH IANEVEENDS IFVKLLKISG IILKTGESQN QLAVDQIAFQ KKLFQTLRR HPSYPKIIEE FVSGLESYIE DEDSFRNCLL SCERLQDEEA SMGASYSKSL IKLLLGIDIL QPAIIKTLFE K LPEYFFEN KNSDEINIPR LIVSQLKWLD RVVDGKDLTT KIMQLISIAP ENLQHDIITS LPEILGDSQH ADVGKELSDL LI ENTSLTV PILDVLSSLR LDPNFLLKVR QLVMDKLSSI RLEDLPVIIK FILHSVTAMD TLEVISELRE KLDLQHCVLP SRL QASQVK LKSKGRASSS GNQESSGQSC IILLFDVIKS AIRYEKTISE AWIKAIENTA SVSEHKVFDL VMLFIIYSTN TQTK KYIDR VLRNKIRSGC IQEQLLQSTF SVHYLVLKDM CSSILSLAQS LLHSLDQSII SFGSLLYKYA FKFFDTYCQQ EVVGA LVTH ICSGNEAEVD TALDVLLELV VLNPSAMMMN AVFVKGILDY LDNISPQQIR KLFYVLSTLA FSKQNEASSH IQDDMH LVI RKQLSSTVFK YKLIGIIGAV TMAGIMAADR SESPSLTQER ANLSDEQCTQ VTSLLQLVHS CSEQSPQASA LYYDEFA NL IQHEKLDPKA LEWVGHTICN DFQDAFVVDS CVVPEGDFPF PVKALYGLEE YDTQDGIAIN LLPLLFSQDF AKDGGPVT S QESGQKLVSP LCLAPYFRLL RLCVERQHNG NLEEIDGLLD CPIFLTDLEP GEKLESMSAK ERSFMCSLIF LTLNWFREI VNAFCQETSP EMKGKVLTRL KHIVELQIIL EKYLAVTPDY VPPLGNFDVE TLDITPHTVT AISAKIRKKG KIERKQKTDG SKTSSSDTL SEEKNSECDP TPSHRGQLNK EFTGKEEKTS LLLHNSHAFF RELDIEVFSI LHCGLVTKFI LDTEMHTEAT E VVQLGPPE LLFLLEDLSQ KLESMLTPPI ARRVPFLKNK GSRNIGFSHL QQRSAQEIVH CVFQLLTPMC NHLENIHNYF QC LAAENHG VVDGPGVKVQ EYHIMSSCYQ RLLQIFHGLF AWSGFSQPEN QNLLYSALHV LSSRLKQGEH SQPLEELLSQ SVH YLQNFH QSIPSFQCAL YLIRLLMVIL EKSTASAQNK EKIASLARQF LCRVWPSGDK EKSNISNDQL HALLCIYLEH TESI LKAIE EIAGVGVPEL INSPKDASSS TFPTLTRHTF VVFFRVMMAE LEKTVKKIEP GTAADSQQIH EEKLLYWNMA VRDFS ILIN LIKVFDSHPV LHVCLKYGRL FVEAFLKQCM PLLDFSFRKH REDVLSLLET FQLDTRLLHH LCGHSKIHQD TRLTQH VPL LKKTLELLVC RVKAMLTLNN CREAFWLGNL KNRDLQGEEI KSQNSQESTA DESEDDMSSQ ASKSKATEDG EEDEVSA GE KEQDSDESYD DSD |
-Macromolecule #3: Polyubiquitin-C
Macromolecule | Name: Polyubiquitin-C / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 8.875125 KDa |
Recombinant expression | Organism: Escherichia coli BL21(DE3) (bacteria) |
Sequence | String: GPGSMQIFVK TLTGKTITLE VEPSDTIENV KAKIQDKEGI PPDQQRLIFA GKQLEDGRTL SDYNIQKEST LHLVLRLRGG |
-Macromolecule #4: Ubiquitin carboxyl-terminal hydrolase 1
Macromolecule | Name: Ubiquitin carboxyl-terminal hydrolase 1 / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO / EC number: ubiquitinyl hydrolase 1 |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 88.390273 KDa |
Recombinant expression | Organism: Spodoptera frugiperda (fall armyworm) |
Sequence | String: GMPGVIPSES NGLSRGSPSK KNRLSLKFFQ KKETKRALDF TDSQENEEKA SEYRASEIDQ VVPAAQSSPI NCEKRENLLP FVGLNNLGN TSYLNSILQV LYFCPGFKSG VKHLFNIISR KKEALKDEAN QKDKGNCKED SLASYELICS LQSLIISVEQ L QASFLLNP ...String: GMPGVIPSES NGLSRGSPSK KNRLSLKFFQ KKETKRALDF TDSQENEEKA SEYRASEIDQ VVPAAQSSPI NCEKRENLLP FVGLNNLGN TSYLNSILQV LYFCPGFKSG VKHLFNIISR KKEALKDEAN QKDKGNCKED SLASYELICS LQSLIISVEQ L QASFLLNP EKYTDELATQ PRRLLNTLRE LNPMYEGYLQ HDAQEVLQCI LGNIQETCQL LKKEEVKNVA ELPTKVEEIP HP KEEMNGI NSIEMDSMRH SEDFKEKLPK GNGKRKSDTE FGNMKKKVKL SKEHQSLEEN QRQTRSKRKA TSDTLESPPK IIP KYISEN ESPRPSQKKS RVKINWLKSA TKQPSILSKF CSLGKITTNQ GVKGQSKENE CDPEEDLGKC ESDNTTNGCG LESP GNTVT PVNVNEVKPI NKGEEQIGFE LVEKLFQGQL VLRTRCLECE SLTERREDFQ DISVPVQEDE LSKVEESSEI SPEPK TEMK TLRWAISQFA SVERIVGEDK YFCENCHHYT EAERSLLFDK MPEVITIHLK CFAASGLEFD CYGGGLSKIN TPLLTP LKL SLEEWSTKPT NDSYGLFAVV MHSGITISSG HYTASVKVTD LNSLELDKGN FVVDQMCEIG KPEPLNEEEA RGVVENY ND EEVSIRVGGN TQPSKVLNKK NVEAIGLLAA QKSKADYELY NKASNPDKVA STAFAENRNS ETSDTTGTHE SDRNKESS D QTGINISGFE NKISYVVQSL KEYEGKWLLF DDSEVKVTEE KDFLNSLSPS TSPTSTPYLL FYKKL |
-Macromolecule #5: WD repeat-containing protein 48
Macromolecule | Name: WD repeat-containing protein 48 / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 78.300656 KDa |
Recombinant expression | Organism: Spodoptera frugiperda (fall armyworm) |
Sequence | String: MHHHHHHLEV LFQGPGSMAA HHRQNTAGRR KVQVSYVIRD EVEKYNRNGV NALQLDPALN RLFTAGRDSI IRIWSVNQHK QDPYIASME HHTDWVNDIV LCCNGKTLIS ASSDTTVKVW NAHKGFCMST LRTHKDYVKA LAYAKDKELV ASAGLDRQIF L WDVNTLTA ...String: MHHHHHHLEV LFQGPGSMAA HHRQNTAGRR KVQVSYVIRD EVEKYNRNGV NALQLDPALN RLFTAGRDSI IRIWSVNQHK QDPYIASME HHTDWVNDIV LCCNGKTLIS ASSDTTVKVW NAHKGFCMST LRTHKDYVKA LAYAKDKELV ASAGLDRQIF L WDVNTLTA LTASNNTVTT SSLSGNKDSI YSLAMNQLGT IIVSGSTEKV LRVWDPRTCA KLMKLKGHTD NVKALLLNRD GT QCLSGSS DGTIRLWSLG QQRCIATYRV HDEGVWALQV NDAFTHVYSG GRDRKIYCTD LRNPDIRVLI CEEKAPVLKM ELD RSADPP PAIWVATTKS TVNKWTLKGI HNFRASGDYD NDCTNPITPL CTQPDQVIKG GASIIQCHIL NDKRHILTKD TNNN VAYWD VLKACKVEDL GKVDFEDEIK KRFKMVYVPN WFSVDLKTGM LTITLDESDC FAAWVSAKDA GFSSPDGSDP KLNLG GLLL QALLEYWPRT HVNPMDEEEN EVNHVNGEQE NRVQKGNGYF QVPPHTPVIF GEAGGRTLFR LLCRDSGGET ESMLLN ETV PQWVIDITVD KNMPKFNKIP FYLQPHASSG AKTLKKDRLS ASDMLQVRKV MEHVYEKIIN LDNESQTTSS SNNEKPG EQ EKEEDIAVLA EEKIELLCQD QVLDPNMDLR TVKHFIWKSG GDLTLHYRQK ST |
-Macromolecule #6: DNA (61-MER)
Macromolecule | Name: DNA (61-MER) / type: dna / ID: 6 Details: Actual sequence: TGATCAGAGGTCATTTGAATTCATGGCTTCGAGCTTCATGTAGAGTCGACGGTGCTGGGAT Number of copies: 2 / Classification: DNA |
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Source (natural) | Organism: synthetic construct (others) |
Molecular weight | Theoretical: 5.17782 KDa |
Sequence | String: (DN)(DN)(DN)(DN)(DN)(DN)(DN)(DN)(DN)(DN) (DN)(DN)(DN)(DN)(DN)(DN)(DN)(DN)(DN)(DN) (DN)(DN)(DN)(DN)(DN)(DN)(DN)(DN)(DN) |
-Macromolecule #7: ZINC ION
Macromolecule | Name: ZINC ION / type: ligand / ID: 7 / Number of copies: 1 / Formula: ZN |
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Molecular weight | Theoretical: 65.409 Da |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 8 |
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Vitrification | Cryogen name: ETHANE / Chamber humidity: 95 % / Chamber temperature: 288 K / Details: blotted for 3.0 secs before plunging. |
Details | 9.3 uM USP1-UAF1, 1.8 uM FANCI-FANCD2Ub, 2.2 uM dsDNA (61 base-pairs), 18 uM ML323 |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K3 (6k x 4k) / Average electron dose: 40.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 3.0 µm / Nominal defocus min: 1.0 µm |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |