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- EMDB-25122: Cryo-EM 3D map of the yeast Rad24-RFC loader bound to DNA and the... -

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Basic information

Entry
Database: EMDB / ID: EMD-25122
TitleCryo-EM 3D map of the yeast Rad24-RFC loader bound to DNA and the open 9-1-1 clamp
Map dataCryo-EM map of the yeast Rad24-RFC loader bound to the 911 clamp and DNA in an open state
Sample
  • Complex: Rad24-RFC-911 clamp-DNA
    • Complex: dsDNADNA
      • DNA: x 2 types
    • Complex: ProteinsProtein
      • Protein or peptide: x 8 types
  • Ligand: x 3 types
Function / homology
Function and homology information


checkpoint clamp complex / meiotic recombination checkpoint signaling / DNA clamp unloading / Rad17 RFC-like complex / Elg1 RFC-like complex / DNA replication factor C complex / Ctf18 RFC-like complex / DNA clamp loader activity / Polymerase switching / nuclease activity ...checkpoint clamp complex / meiotic recombination checkpoint signaling / DNA clamp unloading / Rad17 RFC-like complex / Elg1 RFC-like complex / DNA replication factor C complex / Ctf18 RFC-like complex / DNA clamp loader activity / Polymerase switching / nuclease activity / DNA replication checkpoint signaling / Activation of ATR in response to replication stress / mitotic DNA replication checkpoint signaling / reciprocal meiotic recombination / recombinational repair / mitotic intra-S DNA damage checkpoint signaling / sister chromatid cohesion / mitotic sister chromatid cohesion / leading strand elongation / protein kinase activator activity / Gap-filling DNA repair synthesis and ligation in TC-NER / mitotic G2 DNA damage checkpoint signaling / mismatch repair / mitotic G1 DNA damage checkpoint signaling / meiotic cell cycle / DNA damage checkpoint signaling / condensed nuclear chromosome / cellular response to ionizing radiation / nucleotide-excision repair / DNA-templated DNA replication / double-strand break repair / double-stranded DNA binding / damaged DNA binding / DNA repair / chromatin binding / ATP hydrolysis activity / DNA binding / ATP binding / nucleus / cytosol / cytoplasm
Similarity search - Function
Ddc1 / DNA damage checkpoint control protein Rad17 / Rad17 P-loop domain / Checkpoint protein Rad17/Rad24 / Checkpoint protein Rad17/Rad24, fungi/metazoa / Rad1/Rec1/Rad17 / Rad9/Ddc1 / Repair protein Rad1/Rec1/Rad17 / Replication factor C, C-terminal / Replication factor C C-terminal domain ...Ddc1 / DNA damage checkpoint control protein Rad17 / Rad17 P-loop domain / Checkpoint protein Rad17/Rad24 / Checkpoint protein Rad17/Rad24, fungi/metazoa / Rad1/Rec1/Rad17 / Rad9/Ddc1 / Repair protein Rad1/Rec1/Rad17 / Replication factor C, C-terminal / Replication factor C C-terminal domain / : / DNA polymerase III, delta subunit / DNA polymerase III, clamp loader complex, gamma/delta/delta subunit, C-terminal / : / ATPase family associated with various cellular activities (AAA) / ATPase, AAA-type, core / ATPases associated with a variety of cellular activities / AAA+ ATPase domain / P-loop containing nucleoside triphosphate hydrolase
Similarity search - Domain/homology
: / Checkpoint protein RAD24 / Replication factor C subunit 5 / Replication factor C subunit 3 / Replication factor C subunit 4 / Replication factor C subunit 2 / DNA damage checkpoint control protein RAD17 / DNA damage checkpoint protein 1
Similarity search - Component
Biological speciessynthetic construct (others) / Saccharomyces cerevisiae (brewer's yeast)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.23 Å
AuthorsZheng F / Georgescu R / Yao YN / O'Donnell ME / Li H
Funding support United States, 3 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)GM131754 United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)GM115809 United States
Howard Hughes Medical Institute (HHMI) United States
CitationJournal: Nat Struct Mol Biol / Year: 2022
Title: DNA is loaded through the 9-1-1 DNA checkpoint clamp in the opposite direction of the PCNA clamp.
Authors: Fengwei Zheng / Roxana E Georgescu / Nina Y Yao / Michael E O'Donnell / Huilin Li /
Abstract: The 9-1-1 DNA checkpoint clamp is loaded onto 5'-recessed DNA to activate the DNA damage checkpoint that arrests the cell cycle. The 9-1-1 clamp is a heterotrimeric ring that is loaded in ...The 9-1-1 DNA checkpoint clamp is loaded onto 5'-recessed DNA to activate the DNA damage checkpoint that arrests the cell cycle. The 9-1-1 clamp is a heterotrimeric ring that is loaded in Saccharomyces cerevisiae by Rad24-RFC (hRAD17-RFC), an alternate clamp loader in which Rad24 replaces Rfc1 in the RFC1-5 clamp loader of proliferating cell nuclear antigen (PCNA). The 9-1-1 clamp loading mechanism has been a mystery, because, unlike RFC, which loads PCNA onto a 3'-recessed junction, Rad24-RFC loads the 9-1-1 ring onto a 5'-recessed DNA junction. Here we report two cryo-EM structures of Rad24-RFC-DNA with a closed or 27-Å open 9-1-1 clamp. The structures reveal a completely unexpected mechanism by which a clamp can be loaded onto DNA. Unlike RFC, which encircles DNA, Rad24 binds 5'-DNA on its surface, not inside the loader, and threads the 3' ssDNA overhang into the 9-1-1 clamp from above the ring.
History
DepositionOct 7, 2021-
Header (metadata) releaseMar 23, 2022-
Map releaseMar 23, 2022-
UpdateApr 27, 2022-
Current statusApr 27, 2022Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_25122.png

Downloads & links

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Map

FileDownload / File: emd_25122.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationCryo-EM map of the yeast Rad24-RFC loader bound to the 911 clamp and DNA in an open state
Voxel sizeX=Y=Z: 0.828 Å
Density
Contour LevelBy AUTHOR: 0.08
Minimum - Maximum-0.4495905 - 0.7597728
Average (Standard dev.)0.00035058538 (±0.014939015)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions400400400
Spacing400400400
CellA=B=C: 331.2 Å
α=β=γ: 90.0 °

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Supplemental data

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Sample components

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Entire : Rad24-RFC-911 clamp-DNA

EntireName: Rad24-RFC-911 clamp-DNA
Components
  • Complex: Rad24-RFC-911 clamp-DNA
    • Complex: dsDNADNA
      • DNA: Watson strand
      • DNA: Crick strand
    • Complex: ProteinsProtein
      • Protein or peptide: Checkpoint protein RAD24
      • Protein or peptide: Replication factor C subunit 4
      • Protein or peptide: Replication factor C subunit 3
      • Protein or peptide: Replication factor C subunit 2
      • Protein or peptide: Replication factor C subunit 5
      • Protein or peptide: Mitosis Entry Checkpoint protein MEC3
      • Protein or peptide: DNA damage checkpoint control protein RAD17
      • Protein or peptide: DNA damage checkpoint protein DDC1DNA repair
  • Ligand: PHOSPHOTHIOPHOSPHORIC ACID-ADENYLATE ESTER
  • Ligand: MAGNESIUM ION
  • Ligand: ADENOSINE-5'-DIPHOSPHATE

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Supramolecule #1: Rad24-RFC-911 clamp-DNA

SupramoleculeName: Rad24-RFC-911 clamp-DNA / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#10

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Supramolecule #2: dsDNA

SupramoleculeName: dsDNA / type: complex / ID: 2 / Parent: 1 / Macromolecule list: #9-#10 / Details: synthetic DNA
Source (natural)Organism: synthetic construct (others)

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Supramolecule #3: Proteins

SupramoleculeName: Proteins / type: complex / ID: 3 / Parent: 1 / Macromolecule list: #1-#8
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Recombinant expressionOrganism: Saccharomyces cerevisiae (brewer's yeast)

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Macromolecule #1: Checkpoint protein RAD24

MacromoleculeName: Checkpoint protein RAD24 / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 75.823477 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae (brewer's yeast)
SequenceString: MDSTNLNKRP LLQYSLSSLG SQITKWSSSR PTSPVRKARS TENDFLSKQD TSSILPSIND DGGEQWYEKF KPNCLEQVAI HKRKLKDVQ EALDAMFLPN AKHRILLLSG PSGCSKSTVI KELSKILVPK YRQNSNGTSF RSTPNEHKVT EFRGDCIVND L PQMESFSE ...String:
MDSTNLNKRP LLQYSLSSLG SQITKWSSSR PTSPVRKARS TENDFLSKQD TSSILPSIND DGGEQWYEKF KPNCLEQVAI HKRKLKDVQ EALDAMFLPN AKHRILLLSG PSGCSKSTVI KELSKILVPK YRQNSNGTSF RSTPNEHKVT EFRGDCIVND L PQMESFSE FLKGARYLVM SNLSLILIED LPNVFHIDTR RRFQQLILQW LYSSEPLLPP LVICITECEI PENDNNYRKF GI DYTFSAE TIMNKEILMH PRLKRIKFNP INSTLLKKHL KFICVQNMKM LKEKNKWNKR QEVIDYIAQE TGDIRSAITT LQF WATSSG SLPISTREST ISYFHAIGKV IHGSHSTNND NEMINNLFEN SNNLLSKEDF KLGILENYNT FNKGEFSISD ASSI VDCLS ECDNMNGLPE SNEYGLREVR KTFRNISKQG HNHGTVYFPR EWKVRKLQNS FKVQAEDWLN VSLYKYNAVH SFRNI TLEF GYYAPLIRKC QSYKKKYILY YLKNLPSGSS GPKQTMDKFS DIMKVENGID VVDRIGGPIE ALSVEDGLAP LMDNDS NNC DHLEDQKKER DRRLRMLIDQ YERNVMMAND DLEDEETSFN DDPIVDSDSD NSNNIGNETF GRSDEDESLC EILSQRQ PR KAPVISESLS DSDLEIL

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Macromolecule #2: Replication factor C subunit 4

MacromoleculeName: Replication factor C subunit 4 / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 36.201039 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae (brewer's yeast)
SequenceString: MSKTLSLQLP WVEKYRPQVL SDIVGNKETI DRLQQIAKDG NMPHMIISGM PGIGKTTSVH CLAHELLGRS YADGVLELNA SDDRGIDVV RNQIKHFAQK KLHLPPGKHK IVILDEADSM TAGAQQALRR TMELYSNSTR FAFACNQSNK IIEPLQSRCA I LRYSKLSD ...String:
MSKTLSLQLP WVEKYRPQVL SDIVGNKETI DRLQQIAKDG NMPHMIISGM PGIGKTTSVH CLAHELLGRS YADGVLELNA SDDRGIDVV RNQIKHFAQK KLHLPPGKHK IVILDEADSM TAGAQQALRR TMELYSNSTR FAFACNQSNK IIEPLQSRCA I LRYSKLSD EDVLKRLLQI IKLEDVKYTN DGLEAIIFTA EGDMRQAINN LQSTVAGHGL VNADNVFKIV DSPHPLIVKK ML LASNLED SIQILRTDLW KKGYSSIDIV TTSFRVTKNL AQVKESVRLE MIKEIGLTHM RILEGVGTYL QLASMLAKIH KLN NKA

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Macromolecule #3: Replication factor C subunit 3

MacromoleculeName: Replication factor C subunit 3 / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 38.254543 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae (brewer's yeast)
SequenceString: MSTSTEKRSK ENLPWVEKYR PETLDEVYGQ NEVITTVRKF VDEGKLPHLL FYGPPGTGKT STIVALAREI YGKNYSNMVL ELNASDDRG IDVVRNQIKD FASTRQIFSK GFKLIILDEA DAMTNAAQNA LRRVIERYTK NTRFCVLANY AHKLTPALLS R CTRFRFQP ...String:
MSTSTEKRSK ENLPWVEKYR PETLDEVYGQ NEVITTVRKF VDEGKLPHLL FYGPPGTGKT STIVALAREI YGKNYSNMVL ELNASDDRG IDVVRNQIKD FASTRQIFSK GFKLIILDEA DAMTNAAQNA LRRVIERYTK NTRFCVLANY AHKLTPALLS R CTRFRFQP LPQEAIERRI ANVLVHEKLK LSPNAEKALI ELSNGDMRRV LNVLQSCKAT LDNPDEDEIS DDVIYECCGA PR PSDLKAV LKSILEDDWG TAHYTLNKVR SAKGLALIDL IEGIVKILED YELQNEETRV HLLTKLADIE YSISKGGNDQ IQG SAVIGA IKASFENETV KANV

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Macromolecule #4: Replication factor C subunit 2

MacromoleculeName: Replication factor C subunit 2 / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 39.794473 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae (brewer's yeast)
SequenceString: MFEGFGPNKK RKISKLAAEQ SLAQQPWVEK YRPKNLDEVT AQDHAVTVLK KTLKSANLPH MLFYGPPGTG KTSTILALTK ELYGPDLMK SRILELNASD ERGISIVREK VKNFARLTVS KPSKHDLENY PCPPYKIIIL DEADSMTADA QSALRRTMET Y SGVTRFCL ...String:
MFEGFGPNKK RKISKLAAEQ SLAQQPWVEK YRPKNLDEVT AQDHAVTVLK KTLKSANLPH MLFYGPPGTG KTSTILALTK ELYGPDLMK SRILELNASD ERGISIVREK VKNFARLTVS KPSKHDLENY PCPPYKIIIL DEADSMTADA QSALRRTMET Y SGVTRFCL ICNYVTRIID PLASRCSKFR FKALDASNAI DRLRFISEQE NVKCDDGVLE RILDISAGDL RRGITLLQSA SK GAQYLGD GKNITSTQVE ELAGVVPHDI LIEIVEKVKS GDFDEIKKYV NTFMKSGWSA ASVVNQLHEY YITNDNFDTN FKN QISWLL FTTDSRLNNG TNEHIQLLNL LVKISQL

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Macromolecule #5: Replication factor C subunit 5

MacromoleculeName: Replication factor C subunit 5 / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 39.993582 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae (brewer's yeast)
SequenceString: MSLWVDKYRP KSLNALSHNE ELTNFLKSLS DQPRDLPHLL LYGPNGTGKK TRCMALLESI FGPGVYRLKI DVRQFVTASN RKLELNVVS SPYHLEITPS DMGNNDRIVI QELLKEVAQM EQVDFQDSKD GLAHRYKCVI INEANSLTKD AQAALRRTME K YSKNIRLI ...String:
MSLWVDKYRP KSLNALSHNE ELTNFLKSLS DQPRDLPHLL LYGPNGTGKK TRCMALLESI FGPGVYRLKI DVRQFVTASN RKLELNVVS SPYHLEITPS DMGNNDRIVI QELLKEVAQM EQVDFQDSKD GLAHRYKCVI INEANSLTKD AQAALRRTME K YSKNIRLI MVCDSMSPII APIKSRCLLI RCPAPSDSEI STILSDVVTN ERIQLETKDI LKRIAQASNG NLRVSLLMLE SM ALNNELA LKSSSPIIKP DWIIVIHKLT RKIVKERSVN SLIECRAVLY DLLAHCIPAN IILKELTFSL LDVETLNTTN KSS IIEYSS VFDERLSLGN KAIFHLEGFI AKVMCCLD

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Macromolecule #6: Mitosis Entry Checkpoint protein MEC3

MacromoleculeName: Mitosis Entry Checkpoint protein MEC3 / type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 50.407812 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae (brewer's yeast)
SequenceString: MKLKLIVNGC EAPDDYKLLR TTINTVASLR KTAILRFNSE RLTIISTPKS SLNSSNNGTI LRGDTGQLWC TIPHDVFRLY TVISARELN TITMECNCDS LLSVFKRYDR VMNQGSSSNM TIKLQSCALG ITFEEIVHTS GPNDAIVMNG GVDEHNGLPT T VGTGNLLA ...String:
MKLKLIVNGC EAPDDYKLLR TTINTVASLR KTAILRFNSE RLTIISTPKS SLNSSNNGTI LRGDTGQLWC TIPHDVFRLY TVISARELN TITMECNCDS LLSVFKRYDR VMNQGSSSNM TIKLQSCALG ITFEEIVHTS GPNDAIVMNG GVDEHNGLPT T VGTGNLLA SNKVIMHSFK VPVKLLFRAQ DTRIQEPMIN YIQLMMYKLP PISGEFGSAF HGFIRRVERY SNVNHIHLMG VK KKEHGNE GDDVELKIIV NELDWHLEIC WNGPLDSVIQ RQEGLTDNPS QNQHIDTDGR QEEGSLPIIE ADKPMSSLYT NTR DREMEE NIRYDEDLLR IEDSSIADTR GNIYTADTSG DTEFNDISVM VEKAEQESSS THEVIIRCKD WKVCSKLYAA FEEV VLAIS HDESCVFHCS LDRGSLEDSE DVEKPRERGQ IIYYIARSKG L

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Macromolecule #7: DNA damage checkpoint control protein RAD17

MacromoleculeName: DNA damage checkpoint control protein RAD17 / type: protein_or_peptide / ID: 7 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 45.637527 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae (brewer's yeast)
SequenceString: MRINSELANK FSASTVHLEH ITTALSCLTP FGSKDDVLIF IDADGLSFVR ENNHVIKIQL LLSRELFMSY SYRNETEDHM KLCVKINHI LDSVSVMNRN SDDIVECTLS YDGHGSPFVL IFEDSFISER VEYSTYLIKD FDTNGLELDR ERISFEAIIK G EALHSALK ...String:
MRINSELANK FSASTVHLEH ITTALSCLTP FGSKDDVLIF IDADGLSFVR ENNHVIKIQL LLSRELFMSY SYRNETEDHM KLCVKINHI LDSVSVMNRN SDDIVECTLS YDGHGSPFVL IFEDSFISER VEYSTYLIKD FDTNGLELDR ERISFEAIIK G EALHSALK DLKEIGCKEC YVYAKTEAND ENVFALISKS QLGFSKIKLP SNRSILEKLQ VFDGDSTTVI DGFAVIGFFD FT SFDKIRK STKIASKVLF RMDVHGVLSV NILSQTDDVI ITDTTRPSNN RPGSIRQLQL PKDYPGIVIE VCMLEKESID EAA QTEIEL LMETNELGNR NSFKKSTIRK RYGTDKGNET SNDNLLQLNG KKIKLPSEEE NNKNRESEDE ENHCKYPTKD IPIF F

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Macromolecule #8: DNA damage checkpoint protein DDC1

MacromoleculeName: DNA damage checkpoint protein DDC1 / type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 69.7875 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae (brewer's yeast)
SequenceString: MSFKATITES GKQNIWFRAI YVLSTIQDDI KITVTTNELI AWSMNETDTT LCQVRFQKSF FEEYEFKPHE IVFGENGVQV IEDTYGNSH KLYSFRVNGR HLTTISRKPD GDGIKSFTIA VNNTSTCPES LANRLIVVIE MDSLIVKEYC PQFQPIKYDP I IINLKYKR ...String:
MSFKATITES GKQNIWFRAI YVLSTIQDDI KITVTTNELI AWSMNETDTT LCQVRFQKSF FEEYEFKPHE IVFGENGVQV IEDTYGNSH KLYSFRVNGR HLTTISRKPD GDGIKSFTIA VNNTSTCPES LANRLIVVIE MDSLIVKEYC PQFQPIKYDP I IINLKYKR RFLDVFGTAA SDRNPQEPLD PKLLDVFTNT ERELTSALFN EEVESDIRKR NQLTAADEIN YICCNSTLLK NF LDNCNVN VTDEVKLEIN VHRLSITAFT KAVYGKNNDL LRNALSMSNT ISTLDLEHYC LFTTIEDEKQ DKRSHSKRRE HMK SIIFKL KDFKNFITIG PSWKTTQDGN DNISLWFCHP GDPILMQMQK PGVKLELVEV TDSNINDDIL EGKFIKTAIS GSKE EAGLK DNKESCESPL KSKTALKREN LPHSVAGTRN SPLKVSYLTP DNGSTVAKTY RNNTARKLFV EEQSQSTNYE QDKRF RQAS SVHMNMNREQ SFDIGTTHEV ACPRNESNSL KRSIADICNE TEDPTQQSTF AKRADTTVTW GKALPAADDE VSCSNI DRK GMLKKEKLKH MQGLLNSQND TSNHKKQDNK EMEDGLGLTQ VEKPRGIFD

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Macromolecule #9: Watson strand

MacromoleculeName: Watson strand / type: dna / ID: 9 / Number of copies: 1 / Classification: DNA
Source (natural)Organism: synthetic construct (others)
Molecular weightTheoretical: 6.136008 KDa
SequenceString:
(DG)(DC)(DA)(DG)(DA)(DC)(DA)(DC)(DT)(DA) (DC)(DG)(DA)(DG)(DT)(DA)(DC)(DA)(DT)(DA)

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Macromolecule #10: Crick strand

MacromoleculeName: Crick strand / type: dna / ID: 10 / Number of copies: 1 / Classification: DNA
Source (natural)Organism: synthetic construct (others)
Molecular weightTheoretical: 12.214818 KDa
SequenceString:
(DT)(DT)(DT)(DT)(DT)(DT)(DT)(DT)(DT)(DT) (DT)(DA)(DT)(DG)(DT)(DA)(DC)(DT)(DC)(DG) (DT)(DA)(DG)(DT)(DG)(DT)(DC)(DT)(DG) (DC)(DT)(DT)(DT)(DT)(DT)(DT)(DT)(DT)(DT) (DT)

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Macromolecule #11: PHOSPHOTHIOPHOSPHORIC ACID-ADENYLATE ESTER

MacromoleculeName: PHOSPHOTHIOPHOSPHORIC ACID-ADENYLATE ESTER / type: ligand / ID: 11 / Number of copies: 4 / Formula: AGS
Molecular weightTheoretical: 523.247 Da
Chemical component information

ChemComp-AGS:
PHOSPHOTHIOPHOSPHORIC ACID-ADENYLATE ESTER / ATP-gamma-S, energy-carrying molecule analogue*YM

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Macromolecule #12: MAGNESIUM ION

MacromoleculeName: MAGNESIUM ION / type: ligand / ID: 12 / Number of copies: 4 / Formula: MG
Molecular weightTheoretical: 24.305 Da

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Macromolecule #13: ADENOSINE-5'-DIPHOSPHATE

MacromoleculeName: ADENOSINE-5'-DIPHOSPHATE / type: ligand / ID: 13 / Number of copies: 1 / Formula: ADP
Molecular weightTheoretical: 427.201 Da
Chemical component information

ChemComp-ADP:
ADENOSINE-5'-DIPHOSPHATE / ADP, energy-carrying molecule*YM / Adenosine diphosphate

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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: 7.5
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 K3 (6k x 4k) / Average electron dose: 65.0 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Initial angle assignmentType: NOT APPLICABLE
Final angle assignmentType: NOT APPLICABLE
Final reconstructionResolution.type: BY AUTHOR / Resolution: 3.23 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 302403

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