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- EMDB-24297: State E2 nucleolar 60S ribosomal biogenesis intermediate - L1 sta... -

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Basic information

Entry
Database: EMDB / ID: EMD-24297
TitleState E2 nucleolar 60S ribosomal biogenesis intermediate - L1 stalk local map
Map dataState E2 L1-stalk locally refined map
Sample
  • Complex: Nucleolar 60S intermediate purified with tags on Ytm1 and Spb4
    • RNA: x 1 types
    • Protein or peptide: x 10 types
Keywordsribosome biogenesis / DEAD-box ATPases / methyltransferase / nucleolus / RIBOSOME
Function / homology
Function and homology information


25S rRNA (cytosine2870-C5)-methyltransferase / 27S pre-rRNA (guanosine2922-2'-O)-methyltransferase / rRNA (guanosine-2'-O-)-methyltransferase activity / exonucleolytic trimming to generate mature 5'-end of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / rRNA (uridine-2'-O-)-methyltransferase activity / rRNA (guanine) methyltransferase activity / endonucleolytic cleavage in ITS1 upstream of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / rRNA (cytosine-C5-)-methyltransferase activity / endonucleolytic cleavage in 5'-ETS of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / endonucleolytic cleavage to generate mature 5'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) ...25S rRNA (cytosine2870-C5)-methyltransferase / 27S pre-rRNA (guanosine2922-2'-O)-methyltransferase / rRNA (guanosine-2'-O-)-methyltransferase activity / exonucleolytic trimming to generate mature 5'-end of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / rRNA (uridine-2'-O-)-methyltransferase activity / rRNA (guanine) methyltransferase activity / endonucleolytic cleavage in ITS1 upstream of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / rRNA (cytosine-C5-)-methyltransferase activity / endonucleolytic cleavage in 5'-ETS of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / endonucleolytic cleavage to generate mature 5'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / intracellular mRNA localization / rRNA base methylation / rRNA primary transcript binding / maturation of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / rRNA methylation / protein folding chaperone complex / preribosome, large subunit precursor / ribosomal large subunit export from nucleus / mRNA transport / endonucleolytic cleavage in ITS1 to separate SSU-rRNA from 5.8S rRNA and LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of LSU-rRNA / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / assembly of large subunit precursor of preribosome / ribosomal large subunit biogenesis / rRNA processing / 5S rRNA binding / ribosomal large subunit assembly / negative regulation of translation / ribonucleoprotein complex / mRNA binding / nucleolus / RNA binding / nucleoplasm / identical protein binding / nucleus / cytoplasm
Similarity search - Function
60S ribosomal subunit assembly/export protein Loc1 / Ribosome biogenesis factor, NIP7 / RNA (C5-cytosine) methyltransferase, NOP2 / UPF0113, pre-PUA domain / UPF0113 Pre-PUA domain / UPF0113 PUA domain / UPF0113, PUA domain / Nop2p / Bacterial Fmu (Sun)/eukaryotic nucleolar NOL1/Nop2p, conserved site / NOL1/NOP2/sun family signature. ...60S ribosomal subunit assembly/export protein Loc1 / Ribosome biogenesis factor, NIP7 / RNA (C5-cytosine) methyltransferase, NOP2 / UPF0113, pre-PUA domain / UPF0113 Pre-PUA domain / UPF0113 PUA domain / UPF0113, PUA domain / Nop2p / Bacterial Fmu (Sun)/eukaryotic nucleolar NOL1/Nop2p, conserved site / NOL1/NOP2/sun family signature. / Ribosomal RNA methyltransferase, SPB1-like, C-terminal / Ribosomal RNA methyltransferase Spb1, domain of unknown function DUF3381 / AdoMet-dependent rRNA methyltransferase SPB1-like / Spb1 C-terminal domain / Ribosomal RNA methyltransferase Spb1, DUF3381 / : / Ribosomal RNA large subunit methyltransferase E / Ribosome biogenesis protein BRX1 / SAM-dependent methyltransferase RsmB/NOP2-type / RNA (C5-cytosine) methyltransferase / : / 16S rRNA methyltransferase RsmB/F / SAM-dependent MTase RsmB/NOP-type domain profile. / Putative RNA-binding Domain in PseudoUridine synthase and Archaeosine transglycosylase / PUA domain / PUA domain superfamily / PUA domain profile. / Ribosomal biogenesis NSA2 family / Brix domain / Brix domain / Brix domain profile. / Brix / PUA-like superfamily / Ribosomal RNA methyltransferase, FtsJ domain / FtsJ-like methyltransferase / Ribosomal protein S8e/ribosomal biogenesis NSA2 / Ribosomal protein S8e / S-adenosyl-L-methionine-dependent methyltransferase superfamily
Similarity search - Domain/homology
Ribosome biogenesis protein NSA2 homolog / : / : / 27S pre-rRNA (guanosine(2922)-2'-O)-methyltransferase / 25S rRNA (cytosine(2870)-C(5))-methyltransferase / 60S ribosomal subunit assembly/export protein LOC1 / Ribosome biogenesis protein BRX1 / 60S ribosome subunit biogenesis protein NIP7
Similarity search - Component
Biological speciesSaccharomyces cerevisiae BY4741 (yeast)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.71 Å
AuthorsCruz VE / Sekulski K / Peddada N / Erzberger JP
Funding support United States, 3 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)GM135617-01 United States
Cancer Prevention and Research Institute of Texas (CPRIT)RR150074 United States
Robert A. Welch FoundationI-1897 United States
CitationJournal: Nat Struct Mol Biol / Year: 2022
Title: Sequence-specific remodeling of a topologically complex RNP substrate by Spb4.
Authors: Victor Emmanuel Cruz / Kamil Sekulski / Nagesh Peddada / Carolin Sailer / Sahana Balasubramanian / Christine S Weirich / Florian Stengel / Jan P Erzberger /
Abstract: DEAD-box ATPases are ubiquitous enzymes essential in all aspects of RNA biology. However, the limited in vitro catalytic activities described for these enzymes are at odds with their complex cellular ...DEAD-box ATPases are ubiquitous enzymes essential in all aspects of RNA biology. However, the limited in vitro catalytic activities described for these enzymes are at odds with their complex cellular roles, most notably in driving large-scale RNA remodeling steps during the assembly of ribonucleoproteins (RNPs). We describe cryo-EM structures of 60S ribosomal biogenesis intermediates that reveal how context-specific RNA unwinding by the DEAD-box ATPase Spb4 results in extensive, sequence-specific remodeling of rRNA secondary structure. Multiple cis and trans interactions stabilize Spb4 in a post-catalytic, high-energy intermediate that drives the organization of the three-way junction at the base of rRNA domain IV. This mechanism explains how limited strand separation by DEAD-box ATPases is leveraged to provide non-equilibrium directionality and ensure efficient and accurate RNP assembly.
History
DepositionJun 24, 2021-
Header (metadata) releaseNov 9, 2022-
Map releaseNov 9, 2022-
UpdateJun 5, 2024-
Current statusJun 5, 2024Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_24297.png

Downloads & links

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Map

FileDownload / File: emd_24297.map.gz / Format: CCP4 / Size: 282.6 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationState E2 L1-stalk locally refined map
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
1.08 Å/pix.
x 420 pix.
= 453.6 Å		1.08 Å/pix.
x 420 pix.
= 453.6 Å		1.08 Å/pix.
x 420 pix.
= 453.6 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 1.08 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.015
Minimum - Maximum-0.026723512 - 0.060703777
Average (Standard dev.)0.000014362449 (±0.0011299645)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions420420420
Spacing420420420
CellA=B=C: 453.6 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: State E2 L1-stalk locally refined half map 1

Fileemd_24297_half_map_1.map
AnnotationState E2 L1-stalk locally refined half map 1
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: State E2 L1-stalk locally refined half map 2

Fileemd_24297_half_map_2.map
AnnotationState E2 L1-stalk locally refined half map 2
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Sample components

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Entire : Nucleolar 60S intermediate purified with tags on Ytm1 and Spb4

EntireName: Nucleolar 60S intermediate purified with tags on Ytm1 and Spb4
Components
  • Complex: Nucleolar 60S intermediate purified with tags on Ytm1 and Spb4
    • RNA: 25S rRNA
    • Protein or peptide: 60S ribosome subunit biogenesis protein NIP7
    • Protein or peptide: Ribosome biogenesis protein ERB1
    • Protein or peptide: 25S rRNA (cytosine(2870)-C(5))-methyltransferase
    • Protein or peptide: Ribosome biogenesis protein NSA2 homolog
    • Protein or peptide: 27S pre-rRNA (guanosine(2922)-2'-O)-methyltransferase
    • Protein or peptide: 60S ribosomal subunit assembly/export protein LOC1
    • Protein or peptide: Ribosome biogenesis protein BRX1
    • Protein or peptide: rRNA-processing protein EBP2
    • Protein or peptide: Nucleolar GTP-binding protein 1
    • Protein or peptide: 60S ribosomal protein L1-A

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Supramolecule #1: Nucleolar 60S intermediate purified with tags on Ytm1 and Spb4

SupramoleculeName: Nucleolar 60S intermediate purified with tags on Ytm1 and Spb4
type: complex / ID: 1 / Parent: 0 / Macromolecule list: all
Source (natural)Organism: Saccharomyces cerevisiae BY4741 (yeast)
Molecular weightTheoretical: 3.3 MDa

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Macromolecule #1: 25S rRNA

MacromoleculeName: 25S rRNA / type: rna / ID: 1 / Number of copies: 1
Source (natural)Organism: Saccharomyces cerevisiae BY4741 (yeast) / Strain: BY4741
Molecular weightTheoretical: 72.858875 KDa
SequenceString: GCGGUCGAUC GUUUCCCUCA GGAUAUUUUU GGUAAGCAGA ACUACCCUGU UGAGCUUGAC UCUAGUUUGA CAUUGUGAAG AGACAUAGA GGGUGUAGAA UAAGUGGGAG CUUCGGCGCC AGUGAAAUAC CACUACCUUU AUAGUUUCUU UACUAUUGUC A GGUGGGGA ...String:
GCGGUCGAUC GUUUCCCUCA GGAUAUUUUU GGUAAGCAGA ACUACCCUGU UGAGCUUGAC UCUAGUUUGA CAUUGUGAAG AGACAUAGA GGGUGUAGAA UAAGUGGGAG CUUCGGCGCC AGUGAAAUAC CACUACCUUU AUAGUUUCUU UACUAUUGUC A GGUGGGGA GUAAAGUUAC CACAGGGAUA AUUGUGGCAG UCUUGCUGAC CGUCGUGAGA CAGGUUAG

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Macromolecule #2: 60S ribosome subunit biogenesis protein NIP7

MacromoleculeName: 60S ribosome subunit biogenesis protein NIP7 / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae BY4741 (yeast) / Strain: BY4741
Molecular weightTheoretical: 20.411699 KDa
SequenceString:
MRQLTEEETK VVFEKLAGYI GRNISFLVDN KELPHVFRLQ KDRVYYVPDH VAKLATSVAR PNLMSLGICL GKFTKTGKFR LHITSLTVL AKHAKYKIWI KPNGEMPFLY GNHVLKAHVG KMSDDIPEHA GVIVFAMNDV PLGFGVSAKS TSESRNMQPT G IVAFRQAD IGEYLRDEDT LFT

UniProtKB: 60S ribosome subunit biogenesis protein NIP7

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Macromolecule #3: Ribosome biogenesis protein ERB1

MacromoleculeName: Ribosome biogenesis protein ERB1 / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae BY4741 (yeast) / Strain: BY4741
Molecular weightTheoretical: 6.833629 KDa
SequenceString:
TQNTIGNIPL SAYDEMPHIG YDINGKRIMR PAKGSALDQL LDSIELPEGW TGLLDKNSGS SLN

UniProtKB: UNIPROTKB: A0A6V8S2P3

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Macromolecule #4: 25S rRNA (cytosine(2870)-C(5))-methyltransferase

MacromoleculeName: 25S rRNA (cytosine(2870)-C(5))-methyltransferase / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO / EC number: 25S rRNA (cytosine2870-C5)-methyltransferase
Source (natural)Organism: Saccharomyces cerevisiae BY4741 (yeast) / Strain: BY4741
Molecular weightTheoretical: 69.912164 KDa
SequenceString: MGSRRHKNKQ AAPPTLEEFQ ARKEKKANRK LEKGKRPSTT QGDEVSDRKK KKSKPFKKSR KEEEEVVEED KDLPEVDLEE LSKARKSLF DDEEDDDEAG LVDEELKDEF DLEQEYDYDE DEDNDAHPIF SDDDDEADLE ELNAQNMEAL SKKLDEEEAE E AEEAEMEL ...String:
MGSRRHKNKQ AAPPTLEEFQ ARKEKKANRK LEKGKRPSTT QGDEVSDRKK KKSKPFKKSR KEEEEVVEED KDLPEVDLEE LSKARKSLF DDEEDDDEAG LVDEELKDEF DLEQEYDYDE DEDNDAHPIF SDDDDEADLE ELNAQNMEAL SKKLDEEEAE E AEEAEMEL VEAENMQPRA DILPTEEQEE MMAQETPNLT STRTRMIEIV KVLENFKTLG AEGRSRGEYV DRLLKDICEY FG YTPFLAE KLFNLFSPAE AMEFFEANEI ARPITIRTNT LKTRRRDLAQ TLVNRGVNLQ PIGSWTKVGL QIFDSQVPIG ATP EYLAGH YILQAASSFL PVIALDPHEN ERILDMAAAP GGKTTYISAM MKNTGCVFAN DANKSRTKSL IANIHRLGCT NTIV CNYDA REFPKVIGGF DRILLDAPCS GTGVIGKDQS VKVSRTEKDF IQIPHLQKQL LLSAIDSVDC NSKHGGVIVY STCSV AVEE DEAVIDYALR KRPNVKLVDT GLAIGKEAFT SYRGKKFHPS VKLARRYYPH TYNVDGFFVA KFQKIGPSSF DDNQAS AKE KETAARKEAL EEGIIHSDFA TFEDEEDDKY IEKSVKNNLL KKGVNPKAKR PSNEK

UniProtKB: 25S rRNA (cytosine(2870)-C(5))-methyltransferase

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Macromolecule #5: Ribosome biogenesis protein NSA2 homolog

MacromoleculeName: Ribosome biogenesis protein NSA2 homolog / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae BY4741 (yeast) / Strain: BY4741
Molecular weightTheoretical: 11.673679 KDa
SequenceString:
RMITKHTFVG EGFTRRPVKM ERIIRPSALR QKKANVTHPE LGVTVFLPIL AVKKNPQSPM YTQLGVLTKG TIIEVNVSEL VWGKYAQVT NEPDRDGCVN AVLLD

UniProtKB: Ribosome biogenesis protein NSA2 homolog

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Macromolecule #6: 27S pre-rRNA (guanosine(2922)-2'-O)-methyltransferase

MacromoleculeName: 27S pre-rRNA (guanosine(2922)-2'-O)-methyltransferase / type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO
EC number: 27S pre-rRNA (guanosine2922-2'-O)-methyltransferase
Source (natural)Organism: Saccharomyces cerevisiae BY4741 (yeast) / Strain: BY4741
Molecular weightTheoretical: 1.520849 KDa
SequenceString:
QRALRRIAKK HH

UniProtKB: 27S pre-rRNA (guanosine(2922)-2'-O)-methyltransferase

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Macromolecule #7: 60S ribosomal subunit assembly/export protein LOC1

MacromoleculeName: 60S ribosomal subunit assembly/export protein LOC1 / type: protein_or_peptide / ID: 7 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae BY4741 (yeast) / Strain: BY4741
Molecular weightTheoretical: 23.652645 KDa
SequenceString: MAPKKPSKRQ NLRREVAPEV FQDSQARNQL ANVPHLTEKS AQRKPSKTKV KKEQSLARLY GAKKDKKGKY SEKDLNIPTL NRAIVPGVK IRRGKKGKKF IADNDTLTLN RLITTIGDKY DDIAESKLEK ARRLEEIREL KRKEIERKEA LKQDKLEEKK D EIKKKSSV ...String:
MAPKKPSKRQ NLRREVAPEV FQDSQARNQL ANVPHLTEKS AQRKPSKTKV KKEQSLARLY GAKKDKKGKY SEKDLNIPTL NRAIVPGVK IRRGKKGKKF IADNDTLTLN RLITTIGDKY DDIAESKLEK ARRLEEIREL KRKEIERKEA LKQDKLEEKK D EIKKKSSV ARTIRRKNKR DMLKSEAKAS ESKTEGRKVK KVSFAQ

UniProtKB: 60S ribosomal subunit assembly/export protein LOC1

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Macromolecule #8: Ribosome biogenesis protein BRX1

MacromoleculeName: Ribosome biogenesis protein BRX1 / type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae BY4741 (yeast) / Strain: BY4741
Molecular weightTheoretical: 33.585414 KDa
SequenceString: MSSIYKALAG KSKDNKSEKK QGNVKQFMNK QRTLLISSRG VNYRHRHLIQ DLSGLLPHSR KEPKLDTKKD LQQLNEIAEL YNCNNVLFF EARKHQDLYL WLSKPPNGPT IKFYIQNLHT MDELNFTGNC LKGSRPVLSF DQRFESSPHY QLIKELLVHN F GVPPNARK ...String:
MSSIYKALAG KSKDNKSEKK QGNVKQFMNK QRTLLISSRG VNYRHRHLIQ DLSGLLPHSR KEPKLDTKKD LQQLNEIAEL YNCNNVLFF EARKHQDLYL WLSKPPNGPT IKFYIQNLHT MDELNFTGNC LKGSRPVLSF DQRFESSPHY QLIKELLVHN F GVPPNARK SKPFIDHVMS FSIVDDKIWV RTYEISHSTK NKEEYEDGEE DISLVEIGPR FVMTVILILE GSFGGPKIYE NK QYVSPNV VRAQIKQQAA EEAKSRAEAA VERKIKRREN VLAADPLSND ALFK

UniProtKB: Ribosome biogenesis protein BRX1

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Macromolecule #9: rRNA-processing protein EBP2

MacromoleculeName: rRNA-processing protein EBP2 / type: protein_or_peptide / ID: 9 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae BY4741 (yeast) / Strain: BY4741
Molecular weightTheoretical: 11.4444 KDa
SequenceString:
AFYKQSLDAV LVARDELKRL KVPFKRPLDY FAEMVKSDEH MDKIKGKLIE EASDKKAREE ARRQRQLKKF GKQVQNATLQ KRQLEKRET LEKIKSL

UniProtKB: UNIPROTKB: A0A7I9FH46

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Macromolecule #10: Nucleolar GTP-binding protein 1

MacromoleculeName: Nucleolar GTP-binding protein 1 / type: protein_or_peptide / ID: 10 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae BY4741 (yeast) / Strain: BY4741
Molecular weightTheoretical: 6.401858 KDa
SequenceString:
KITRIRAFYM RKVKSLVEQV ARDYVRLLKF GQSLFQCKQL KRAALGRMAT IVKK

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Macromolecule #11: 60S ribosomal protein L1-A

MacromoleculeName: 60S ribosomal protein L1-A / type: protein_or_peptide / ID: 11 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae BY4741 (yeast) / Strain: BY4741
Molecular weightTheoretical: 24.538826 KDa
SequenceString: MSKITSSQVR EHVKELLKYS NETKKRNFLE TVELQVGLKN YDPQRDKRFS GSLKLPNCPR PNMSICIFGD AFDVDRAKSC GVDAMSVDD LKKLNKNKKL IKKLSKKYNA FIASEVLIKQ VPRLLGPQLS KAGKFPTPVS HNDDLYGKVT DVRSTIKFQL K KVLCLAVA ...String:
MSKITSSQVR EHVKELLKYS NETKKRNFLE TVELQVGLKN YDPQRDKRFS GSLKLPNCPR PNMSICIFGD AFDVDRAKSC GVDAMSVDD LKKLNKNKKL IKKLSKKYNA FIASEVLIKQ VPRLLGPQLS KAGKFPTPVS HNDDLYGKVT DVRSTIKFQL K KVLCLAVA VGNVEMEEDV LVQQILMSVN FFVSLLKKNW QNVGSLVVKS SMGPAFRLY

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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

Concentration0.45 mg/mL
BufferpH: 8
Component:
ConcentrationFormulaName
50.0 mMCH2[CH2NHC(CH2OH)3]2Bis-Tris-Propane
150.0 mMNaClSodium Chloride
10.0 mMMgCl2Magnesium Chloride
1.0 mMC9H15O6PHClTCEP
0.01 % (w/v)NP-40
GridModel: Quantifoil R2/2 / Material: COPPER / Mesh: 300 / Support film - Material: CARBON / Support film - topology: CONTINUOUS / Support film - Film thickness: 0.3
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Image recordingFilm or detector model: GATAN K3 (6k x 4k) / Number grids imaged: 1 / Number real images: 4523 / Average exposure time: 0.05 sec. / Average electron dose: 1.2 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsC2 aperture diameter: 70.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.2 µm / Nominal defocus min: 0.9 µm / Nominal magnification: 81000
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Particle selectionNumber selected: 825096
Startup modelType of model: EMDB MAP
EMDB ID:

3891

Final reconstructionNumber classes used: 1 / Applied symmetry - Point group: C1 (asymmetric) / Algorithm: FOURIER SPACE / Resolution.type: BY AUTHOR / Resolution: 3.71 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION / Number images used: 198000
Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION
Final 3D classificationNumber classes: 4 / Avg.num./class: 118000 / Software - Name: RELION
FSC plot (resolution estimation)

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