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- EMDB-37552: Cryo- EM structure of Mycobacterium smegmatis 50S ribosomal subun... -

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Basic information

Entry
Database: EMDB / ID: EMD-37552
TitleCryo- EM structure of Mycobacterium smegmatis 50S ribosomal subunit (body 1) of 70S ribosome and RafH.
Map dataMycobacterium smegmatis 50S ribosomal subunit (body 1) of 70S ribosome and RafH complex
Sample
  • Complex: 70S ribosome + RafH protein
    • Protein or peptide: x 26 types
    • RNA: x 2 types
KeywordsRibosome / protein synthesis / Mycobacterium smegmatis / hibernation promotion factor / RafH / hypoxia stress / Cryo- EM / Single particle reconstruction
Function / homology
Function and homology information


large ribosomal subunit / transferase activity / tRNA binding / rRNA binding / ribosome / structural constituent of ribosome / ribonucleoprotein complex / translation / metal ion binding / cytoplasm
Similarity search - Function
Ribosomal protein L31 type A / Ribosomal protein L31 signature. / Ribosomal protein L31 / Ribosomal protein L31 superfamily / Ribosomal protein L31 / Ribosomal protein L21, conserved site / Ribosomal protein L21 signature. / : / Ribosomal protein L6, conserved site / Ribosomal protein L6 signature 1. ...Ribosomal protein L31 type A / Ribosomal protein L31 signature. / Ribosomal protein L31 / Ribosomal protein L31 superfamily / Ribosomal protein L31 / Ribosomal protein L21, conserved site / Ribosomal protein L21 signature. / : / Ribosomal protein L6, conserved site / Ribosomal protein L6 signature 1. / Ribosomal protein L17 signature. / Ribosomal protein L9 signature. / Ribosomal protein L9, bacteria/chloroplast / Ribosomal protein L9, C-terminal / Ribosomal protein L9, C-terminal domain / Ribosomal protein L9, C-terminal domain superfamily / Ribosomal protein L28/L24 superfamily / Ribosomal protein L9, N-terminal domain superfamily / Ribosomal protein L9 / Ribosomal protein L9, N-terminal / Ribosomal protein L9, N-terminal domain / Ribosomal protein L28 / Ribosomal protein L35, conserved site / Ribosomal protein L35 signature. / Ribosomal protein L33, conserved site / Ribosomal protein L33 signature. / Ribosomal protein L35, non-mitochondrial / Ribosomal protein L5, bacterial-type / Ribosomal protein L6, bacterial-type / Ribosomal protein L18, bacterial-type / Ribosomal protein L19, conserved site / Ribosomal protein L19 signature. / Ribosomal protein L9/RNase H1, N-terminal / Ribosomal protein L20 signature. / Ribosomal protein L27, conserved site / Ribosomal protein L27 signature. / Ribosomal protein L14P, bacterial-type / Ribosomal protein L34, conserved site / Ribosomal protein L34 signature. / Ribosomal protein L22, bacterial/chloroplast-type / Ribosomal protein L35 / Ribosomal protein L35 superfamily / Ribosomal protein L2, bacterial/organellar-type / Ribosomal protein L35 / Ribosomal L28 family / Ribosomal protein L33 / Ribosomal protein L33 / Ribosomal protein L28/L24 / Ribosomal protein L33 superfamily / : / Ribosomal protein L30, bacterial-type / Ribosomal protein L18 / Ribosomal L18 of archaea, bacteria, mitoch. and chloroplast / L28p-like / Ribosomal protein L20 / Ribosomal protein L20 / Ribosomal protein L20, C-terminal / Ribosomal protein L21 / Ribosomal protein L27 / Ribosomal L27 protein / Ribosomal protein L19 / Ribosomal protein L19 superfamily / Ribosomal protein L19 / Ribosomal proteins 50S L24/mitochondrial 39S L24 / Ribosomal protein L17 / Ribosomal protein L17 superfamily / Ribosomal protein L17 / Ribosomal protein L21-like / L21-like superfamily / Ribosomal prokaryotic L21 protein / Ribosomal L32p protein family / Ribosomal protein L24 / Ribosomal protein L32p / Ribosomal protein L34 / Ribosomal protein L34 / Ribosomal protein L13, bacterial-type / Ribosomal protein L23/L25, conserved site / Ribosomal protein L3, bacterial/organelle-type / Ribosomal protein L15, bacterial-type / 50S ribosomal protein uL4 / Ribosomal protein L23 signature. / Ribosomal protein L30, conserved site / Ribosomal protein L29, conserved site / Ribosomal protein L30 signature. / Ribosomal protein L5, N-terminal / Ribosomal protein L15, conserved site / Ribosomal protein L5, C-terminal / Ribosomal protein L5 / Ribosomal protein L5 domain superfamily / Ribosomal protein L2, conserved site / Ribosomal protein L2 signature. / Ribosomal protein L29 signature. / Ribosomal protein L5 / Ribosomal protein L6, alpha-beta domain / Ribosomal protein L6 / ribosomal L5P family C-terminus / Ribosomal protein L6, alpha-beta domain superfamily / Ribosomal protein L6 / Ribosomal protein L2, domain 3 / Ribosomal protein L14P, conserved site
Similarity search - Domain/homology
Large ribosomal subunit protein bL33A / Large ribosomal subunit protein uL3 / Large ribosomal subunit protein uL4 / Large ribosomal subunit protein uL23 / Large ribosomal subunit protein uL2 / Large ribosomal subunit protein uL22 / Large ribosomal subunit protein uL29 / Large ribosomal subunit protein uL14 / Large ribosomal subunit protein uL24 / Large ribosomal subunit protein uL5 ...Large ribosomal subunit protein bL33A / Large ribosomal subunit protein uL3 / Large ribosomal subunit protein uL4 / Large ribosomal subunit protein uL23 / Large ribosomal subunit protein uL2 / Large ribosomal subunit protein uL22 / Large ribosomal subunit protein uL29 / Large ribosomal subunit protein uL14 / Large ribosomal subunit protein uL24 / Large ribosomal subunit protein uL5 / Large ribosomal subunit protein uL6 / Large ribosomal subunit protein uL18 / Large ribosomal subunit protein uL30 / Large ribosomal subunit protein uL15 / Large ribosomal subunit protein bL17 / Large ribosomal subunit protein uL13 / Large ribosomal subunit protein bL28 / Large ribosomal subunit protein bL19 / Large ribosomal subunit protein bL20 / Large ribosomal subunit protein bL35 / Large ribosomal subunit protein bL27 / Large ribosomal subunit protein bL21 / Large ribosomal subunit protein bL31 / Large ribosomal subunit protein bL32 / Large ribosomal subunit protein bL9 / Large ribosomal subunit protein bL34
Similarity search - Component
Biological speciesMycolicibacterium smegmatis MC2 155 (bacteria)
Methodsingle particle reconstruction / cryo EM / Resolution: 2.7 Å
AuthorsKumar N / Sharma S / Kaushal PS
Funding support India, 1 items
OrganizationGrant numberCountry
Science and Engineering Research Board (SERB) India
CitationJournal: Nat Commun / Year: 2024
Title: Cryo- EM structure of the mycobacterial 70S ribosome in complex with ribosome hibernation promotion factor RafH.
Authors: Niraj Kumar / Shivani Sharma / Prem S Kaushal /
Abstract: Ribosome hibernation is a key survival strategy bacteria adopt under environmental stress, where a protein, hibernation promotion factor (HPF), transitorily inactivates the ribosome. Mycobacterium ...Ribosome hibernation is a key survival strategy bacteria adopt under environmental stress, where a protein, hibernation promotion factor (HPF), transitorily inactivates the ribosome. Mycobacterium tuberculosis encounters hypoxia (low oxygen) as a major stress in the host macrophages, and upregulates the expression of RafH protein, which is crucial for its survival. The RafH, a dual domain HPF, an orthologue of bacterial long HPF (HPF), hibernates ribosome in 70S monosome form, whereas in other bacteria, the HPF induces 70S ribosome dimerization and hibernates its ribosome in 100S disome form. Here, we report the cryo- EM structure of M. smegmatis, a close homolog of M. tuberculosis, 70S ribosome in complex with the RafH factor at an overall 2.8 Å resolution. The N- terminus domain (NTD) of RafH binds to the decoding center, similarly to HPF NTD. In contrast, the C- terminus domain (CTD) of RafH, which is larger than the HPF CTD, binds to a distinct site at the platform binding center of the ribosomal small subunit. The two domain-connecting linker regions, which remain mostly disordered in earlier reported HPF structures, interact mainly with the anti-Shine Dalgarno sequence of the 16S rRNA.
History
DepositionSep 23, 2023-
Header (metadata) releaseFeb 28, 2024-
Map releaseFeb 28, 2024-
UpdateFeb 28, 2024-
Current statusFeb 28, 2024Processing site: PDBj / Status: Released

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Structure visualization

Supplemental images

emd_37552.png

Downloads & links

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Map

FileDownload / File: emd_37552.map.gz / Format: CCP4 / Size: 209.3 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationMycobacterium smegmatis 50S ribosomal subunit (body 1) of 70S ribosome and RafH complex
Voxel sizeX=Y=Z: 1.07 Å
Density
Contour LevelBy AUTHOR: 0.045
Minimum - Maximum-0.04162318 - 0.2797405
Average (Standard dev.)0.002174491 (±0.011432372)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions380380380
Spacing380380380
CellA=B=C: 406.6 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: Mycobacterium smegmatis 50S ribosomal subunit (half 1, body...

Fileemd_37552_half_map_1.map
AnnotationMycobacterium smegmatis 50S ribosomal subunit (half 1, body 1) of 70S ribosome and RafH complex
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: Mycobacterium smegmatis 50S ribosomal subunit (half 2, body...

Fileemd_37552_half_map_2.map
AnnotationMycobacterium smegmatis 50S ribosomal subunit (half 2, body 1) of 70S ribosome and RafH complex
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Sample components

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Entire : 70S ribosome + RafH protein

EntireName: 70S ribosome + RafH protein
Components
  • Complex: 70S ribosome + RafH protein
    • Protein or peptide: 50S ribosomal protein L2
    • Protein or peptide: 50S ribosomal protein L3
    • Protein or peptide: 50S ribosomal protein L4
    • Protein or peptide: 50S ribosomal protein L5
    • Protein or peptide: 50S ribosomal protein L6
    • Protein or peptide: 50S ribosomal protein L9
    • Protein or peptide: 50S ribosomal protein L13
    • Protein or peptide: 50S ribosomal protein L14
    • Protein or peptide: 50S ribosomal protein L15
    • Protein or peptide: 50S ribosomal protein L17
    • Protein or peptide: 50S ribosomal protein L18
    • Protein or peptide: 50S ribosomal protein L19
    • Protein or peptide: 50S ribosomal protein L20
    • Protein or peptide: 50S ribosomal protein L21
    • Protein or peptide: 50S ribosomal protein L22
    • Protein or peptide: 50S ribosomal protein L23
    • Protein or peptide: 50S ribosomal protein L24
    • Protein or peptide: 50S ribosomal protein L27
    • Protein or peptide: 50S ribosomal protein L28
    • Protein or peptide: 50S ribosomal protein L29
    • Protein or peptide: 50S ribosomal protein L30
    • Protein or peptide: 50S ribosomal protein L32
    • Protein or peptide: 50S ribosomal protein L33ARibosome
    • Protein or peptide: 50S ribosomal protein L34
    • Protein or peptide: 50S ribosomal protein L35
    • Protein or peptide: 50S ribosomal protein L31
    • RNA: 23S rRNA23S ribosomal RNA
    • RNA: 5S rRNA5S ribosomal RNA

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Supramolecule #1: 70S ribosome + RafH protein

SupramoleculeName: 70S ribosome + RafH protein / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria) / Strain: mc(2)155

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Macromolecule #1: 50S ribosomal protein L2

MacromoleculeName: 50S ribosomal protein L2 / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria) / Strain: mc(2)155
Molecular weightTheoretical: 30.425945 KDa
SequenceString: MGIRKYKPTT PGRRGASVSD FAEITRSTPE KSLVRPLHGK GGRNAHGRIT TRHKGGGHKR AYRVIDFRRH DKDGVNAKVA HIEYDPNRT ANIALLHYLD GEKRYIIAPQ GLKQGDVIES GANADIKPGN NLPLRNIPAG TVIHAVELRP GGGAKLARSA G VSIQLLGK ...String:
MGIRKYKPTT PGRRGASVSD FAEITRSTPE KSLVRPLHGK GGRNAHGRIT TRHKGGGHKR AYRVIDFRRH DKDGVNAKVA HIEYDPNRT ANIALLHYLD GEKRYIIAPQ GLKQGDVIES GANADIKPGN NLPLRNIPAG TVIHAVELRP GGGAKLARSA G VSIQLLGK EGTYAALRMP SGEIRRVDVR CRATVGEVGN AEQSNINWGK AGRMRWKGKR PTVRGVVMNP VDHPHGGGEG KT SGGRHPV SPWGKPEGRT RKPNKPSDKL IVRRRRTGKK R

UniProtKB: Large ribosomal subunit protein uL2

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Macromolecule #2: 50S ribosomal protein L3

MacromoleculeName: 50S ribosomal protein L3 / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria) / Strain: mc(2)155
Molecular weightTheoretical: 23.011393 KDa
SequenceString: MARKGILGTK LGMTQVFDEN NKVVPVTVVK AGPNVVTRIR TTERDGYSAV QLAYGEISPR KVIKPVAGQF AAAGVNPRRH VAELRLDDE AAVAEYEVGQ ELTAEIFSDG AYVDVTGTSK GKGFAGTMKR HGFRGQGAAH GAQAVHRRPG SIGGCATPGR V FKGTRMSG ...String:
MARKGILGTK LGMTQVFDEN NKVVPVTVVK AGPNVVTRIR TTERDGYSAV QLAYGEISPR KVIKPVAGQF AAAGVNPRRH VAELRLDDE AAVAEYEVGQ ELTAEIFSDG AYVDVTGTSK GKGFAGTMKR HGFRGQGAAH GAQAVHRRPG SIGGCATPGR V FKGTRMSG RMGNDRVTTQ NLKVHKVDAE NGVLLIKGAI PGRNGGLVVV RSAIKRGEK

UniProtKB: Large ribosomal subunit protein uL3

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Macromolecule #3: 50S ribosomal protein L4

MacromoleculeName: 50S ribosomal protein L4 / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria) / Strain: mc(2)155
Molecular weightTheoretical: 22.920949 KDa
SequenceString: MTLKVDVKTP AGKTDGSVEL PAELFDVEPN IALMHQVVTA QLAAKRQGTH STKTRGEVSG GGKKPYRQKG TGRARQGSTR APQFTGGGT VHGPKPRDYS QRTPKKMIAA ALRGALSDRA RNDRIHAVTE LVEGQTPSTK SAKTFLGTLT ENKKVLVVIG R TDEVGAKS ...String:
MTLKVDVKTP AGKTDGSVEL PAELFDVEPN IALMHQVVTA QLAAKRQGTH STKTRGEVSG GGKKPYRQKG TGRARQGSTR APQFTGGGT VHGPKPRDYS QRTPKKMIAA ALRGALSDRA RNDRIHAVTE LVEGQTPSTK SAKTFLGTLT ENKKVLVVIG R TDEVGAKS VRNLPGVHVI SPDQLNTYDV LNADDVVFSV EALNAYISAN SKEGASV

UniProtKB: Large ribosomal subunit protein uL4

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Macromolecule #4: 50S ribosomal protein L5

MacromoleculeName: 50S ribosomal protein L5 / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria) / Strain: mc(2)155
Molecular weightTheoretical: 21.152328 KDa
SequenceString:
MTTTEKALPR LKQRYREEIR EALQQEFNYA NVMQIPGVVK VVVNMGVGDA ARDAKLINGA INDLALITGQ KPEVRRARKS IAQFKLREG MPIGARVTLR GDRMWEFLDR LISIALPRIR DFRGLSPKQF DGTGNYTFGL NEQSMFHEID VDSIDRPRGM D ITVVTTAT NDAEGRALLR ALGFPFKEN

UniProtKB: Large ribosomal subunit protein uL5

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Macromolecule #5: 50S ribosomal protein L6

MacromoleculeName: 50S ribosomal protein L6 / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria) / Strain: mc(2)155
Molecular weightTheoretical: 19.483361 KDa
SequenceString:
MSRIGKQPVP VPSGVDVTIN GQNLSVKGPK GTLTLDVAEP ISVSRAEDGA IVVTRPDDER RSRSLHGLSR TLIANLVTGV TEGYTQKME IFGVGYRVQL KGQNLEFALG YSHPVLIEAP EGITFAVESP TKFSVSGIDK QKVGQISAVI RRLRRPDPYK G KGVRYEGE QIRRKVGKTG K

UniProtKB: Large ribosomal subunit protein uL6

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Macromolecule #6: 50S ribosomal protein L9

MacromoleculeName: 50S ribosomal protein L9 / type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria) / Strain: mc(2)155
Molecular weightTheoretical: 15.949253 KDa
SequenceString:
MKLILTAEVE HLGAAGDTVE VKDGYGRNYL LPRGLAIVAS RGAERQAEEI RRARESKVIR DIEHANELKT ALEGLGDVTL SVNAAGDTG KLFGSVTAAD VVNAIKKAGG PNLDKRTVQL AKAHIKSVGT HPVTVKLHTG VEAKVSLNVV AQ

UniProtKB: Large ribosomal subunit protein bL9

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Macromolecule #7: 50S ribosomal protein L13

MacromoleculeName: 50S ribosomal protein L13 / type: protein_or_peptide / ID: 7 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria) / Strain: mc(2)155
Molecular weightTheoretical: 16.146689 KDa
SequenceString:
MPTYTPKAGD TTRSWYVIDA SDVVLGRLAS AAATLLRGKH KPTFTPNVDG GDFVIVINAD KIAVSGDKLT KKFAYRHSGY PGGLRKRTI GELLEKHPTR VVENAIIGML PHNKLGRQIQ KKLKVYAGPD HPHAAQQPIP FEIKQVAQ

UniProtKB: Large ribosomal subunit protein uL13

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Macromolecule #8: 50S ribosomal protein L14

MacromoleculeName: 50S ribosomal protein L14 / type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria) / Strain: mc(2)155
Molecular weightTheoretical: 13.400625 KDa
SequenceString:
MIQQESRLKV ADNTGAKEIL CIRVLGGSSR RYAGIGDVIV ATVKDAIPGG NVKRGDVVKA VVVRTVKERR RADGSYIKFD ENAAVIIKN DNDPRGTRIF GPVGRELREK KFMKIVSLAP EVL

UniProtKB: Large ribosomal subunit protein uL14

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Macromolecule #9: 50S ribosomal protein L15

MacromoleculeName: 50S ribosomal protein L15 / type: protein_or_peptide / ID: 9 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria) / Strain: mc(2)155
Molecular weightTheoretical: 15.602244 KDa
SequenceString:
MSVIKLHDLK PAPGEKKAKT RVGRGEGSKG KTAGRGTKGT KARKNVPVMF EGGQMPIHMR LPKLKGFKNR FRTEYQVVNV GDINKAFPQ GGTVGVDELV AKGLVRKNSL VKVLGDGKLT VKVDVTANKF SGSAREAITA AGGSATEL

UniProtKB: Large ribosomal subunit protein uL15

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Macromolecule #10: 50S ribosomal protein L17

MacromoleculeName: 50S ribosomal protein L17 / type: protein_or_peptide / ID: 10 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria) / Strain: mc(2)155
Molecular weightTheoretical: 21.892246 KDa
SequenceString: MPKPTKGPRL GGSSSHQSAL LANLATSLFE HGRIKTTEPK ARALRPYAEK LITHAKKGAL HNRREVMKKI RDKDVVHTLF AEIGPFYAD RNGGYTRIIK VENRKGDNAP MAVIELVREK TVTDEANRAR RAAASQAKAD ERADEKADEK AEETVEETTE A PAEESTEA ...String:
MPKPTKGPRL GGSSSHQSAL LANLATSLFE HGRIKTTEPK ARALRPYAEK LITHAKKGAL HNRREVMKKI RDKDVVHTLF AEIGPFYAD RNGGYTRIIK VENRKGDNAP MAVIELVREK TVTDEANRAR RAAASQAKAD ERADEKADEK AEETVEETTE A PAEESTEA AAEETVEETT EAPAEESTEA AEESEAKDDT K

UniProtKB: Large ribosomal subunit protein bL17

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Macromolecule #11: 50S ribosomal protein L18

MacromoleculeName: 50S ribosomal protein L18 / type: protein_or_peptide / ID: 11 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria) / Strain: mc(2)155
Molecular weightTheoretical: 13.711728 KDa
SequenceString:
MAHKPVGQNI SEVRRNARLR RHARLRKKVA GTAEVPRLVV NRSARHIHVQ LVNDLNGTTL AAASSIEADV RAIDGDKKAH SVRVGQLIA ERAKAAGVET VVFDRGGYTY GGRIAALADA AREAGLKF

UniProtKB: Large ribosomal subunit protein uL18

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Macromolecule #12: 50S ribosomal protein L19

MacromoleculeName: 50S ribosomal protein L19 / type: protein_or_peptide / ID: 12 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria) / Strain: mc(2)155
Molecular weightTheoretical: 12.892806 KDa
SequenceString:
MNTLDFVDQA SLRDDIPTFS PGDTVNVHVK VIEGSKERIQ VFKGVVIRRQ GGGISETFTV RKESYGVGVE RTFPVHSPNI DHIDVLTRG DVRRAKLYYL RELRGKKAKI KEKR

UniProtKB: Large ribosomal subunit protein bL19

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Macromolecule #13: 50S ribosomal protein L20

MacromoleculeName: 50S ribosomal protein L20 / type: protein_or_peptide / ID: 13 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria) / Strain: mc(2)155
Molecular weightTheoretical: 14.494693 KDa
SequenceString:
MARVKRALNA QKKRRTVLKA SKGYRGQRSR LYRKAKEQQL HSLTYAYRDR RARKGEFRKL WISRINAAAR ANDITYNRLI QGLKAAGVE VDRKNLAELA VSDPAAFTAL VDVARAALPE DVNAPSGEAA

UniProtKB: Large ribosomal subunit protein bL20

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Macromolecule #14: 50S ribosomal protein L21

MacromoleculeName: 50S ribosomal protein L21 / type: protein_or_peptide / ID: 14 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria) / Strain: mc(2)155
Molecular weightTheoretical: 11.055894 KDa
SequenceString:
MATYAIVKTG GKQYKVAAGD VVKVEKLDSE PGASVSLPVA LVVDGANVTS KADDLAKVAV TAEVLEHTKG PKIRIHKFKN KTGYHKRQG HRQQLTVLKV TGIK

UniProtKB: Large ribosomal subunit protein bL21

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Macromolecule #15: 50S ribosomal protein L22

MacromoleculeName: 50S ribosomal protein L22 / type: protein_or_peptide / ID: 15 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria) / Strain: mc(2)155
Molecular weightTheoretical: 16.3536 KDa
SequenceString:
MSTVTEFPSA TAKARYVRVS ATKARRVIDL VRGKSVEEAL DILRWAPQAA SEPVAKVIAS AAANAQNNEG LDPSTLVVAT VYADEGPTA KRIRPRAQGR AFRIRKRTSH ITVIVESRPP KQKGASAASA RSRRAQGSKA AATKKSAETK EGSE

UniProtKB: Large ribosomal subunit protein uL22

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Macromolecule #16: 50S ribosomal protein L23

MacromoleculeName: 50S ribosomal protein L23 / type: protein_or_peptide / ID: 16 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria) / Strain: mc(2)155
Molecular weightTheoretical: 11.047836 KDa
SequenceString:
MATITDPRDI ILAPVISEKS YGLIEDNVYT FVVHPDSNKT QIKIAIEKIF DVKVDSVNTA NRQGKRKRTR TGFGKRKSTK RAIVKLAAG SKPIDLFGAP A

UniProtKB: Large ribosomal subunit protein uL23

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Macromolecule #17: 50S ribosomal protein L24

MacromoleculeName: 50S ribosomal protein L24 / type: protein_or_peptide / ID: 17 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria) / Strain: mc(2)155
Molecular weightTheoretical: 11.228946 KDa
SequenceString:
MKVHKGDTVL VISGKDKGAK GKVLVAYPDR NKVLVEGVNR IKKHTAVSAN ERGASSGGIV TQEAPIHVSN VMVVDSDGKP TRVGYRIDD ETGKKVRIAK TNGKDI

UniProtKB: Large ribosomal subunit protein uL24

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Macromolecule #18: 50S ribosomal protein L27

MacromoleculeName: 50S ribosomal protein L27 / type: protein_or_peptide / ID: 18 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria) / Strain: mc(2)155
Molecular weightTheoretical: 9.249582 KDa
SequenceString:
MAHKKGASSS RNGRDSAAQR LGVKRFGGQV VKAGEILVRQ RGTHFHPGVN VGRGGDDTLF ALAPGAVEFG AKRGRKTVNI VPVARPEA

UniProtKB: Large ribosomal subunit protein bL27

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Macromolecule #19: 50S ribosomal protein L28

MacromoleculeName: 50S ribosomal protein L28 / type: protein_or_peptide / ID: 19 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria) / Strain: mc(2)155
Molecular weightTheoretical: 6.891016 KDa
SequenceString:
MAAVCDICGK GPGFGKSVSH SHRRTSRRWN PNIQPVRAVT RPGGNKQRIN ACTSCIKAGK VSRA

UniProtKB: Large ribosomal subunit protein bL28

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Macromolecule #20: 50S ribosomal protein L29

MacromoleculeName: 50S ribosomal protein L29 / type: protein_or_peptide / ID: 20 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria) / Strain: mc(2)155
Molecular weightTheoretical: 8.790895 KDa
SequenceString:
MAVGTTPGEL RELTDDELKD KLRESKEELF NLRFQMATGQ LSNNRRLRTV RQEIARVYTV LRERELGLAS GPAGEES

UniProtKB: Large ribosomal subunit protein uL29

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Macromolecule #21: 50S ribosomal protein L30

MacromoleculeName: 50S ribosomal protein L30 / type: protein_or_peptide / ID: 21 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria) / Strain: mc(2)155
Molecular weightTheoretical: 7.022147 KDa
SequenceString:
MAELKITQVR STIGARWKQR ESLRTLGLKK IRQSVVREDN AQTRGLINTV HHLVEVEEVG K

UniProtKB: Large ribosomal subunit protein uL30

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Macromolecule #22: 50S ribosomal protein L32

MacromoleculeName: 50S ribosomal protein L32 / type: protein_or_peptide / ID: 22 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria) / Strain: mc(2)155
Molecular weightTheoretical: 6.467719 KDa
SequenceString:
MAVPKRRMSR ANTRSRRAQW KAEAPGLVTV SVAGQQRKVP RRLLKAARLG LVDLDKR

UniProtKB: Large ribosomal subunit protein bL32

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Macromolecule #23: 50S ribosomal protein L33A

MacromoleculeName: 50S ribosomal protein L33A / type: protein_or_peptide / ID: 23 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria) / Strain: mc(2)155
Molecular weightTheoretical: 6.468501 KDa
SequenceString:
MASSTDVRPK ITLACEVCKH RNYITKKNRR NDPDRLEIKK FCPNCGTHQP HKESR

UniProtKB: Large ribosomal subunit protein bL33A

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Macromolecule #24: 50S ribosomal protein L34

MacromoleculeName: 50S ribosomal protein L34 / type: protein_or_peptide / ID: 24 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria) / Strain: mc(2)155
Molecular weightTheoretical: 5.52258 KDa
SequenceString:
MAKGKRTFQP NNRRRARVHG FRLRMRTRAG RAIVANRRSK GRRALTA

UniProtKB: Large ribosomal subunit protein bL34

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Macromolecule #25: 50S ribosomal protein L35

MacromoleculeName: 50S ribosomal protein L35 / type: protein_or_peptide / ID: 25 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria) / Strain: mc(2)155
Molecular weightTheoretical: 7.298505 KDa
SequenceString:
MPKAKTHSGA SKRFRRTGTG KIVRQKANRR HLLEHKPTKR TRRLDGRTTV SAADNSRINK LLNG

UniProtKB: Large ribosomal subunit protein bL35

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Macromolecule #26: 50S ribosomal protein L31

MacromoleculeName: 50S ribosomal protein L31 / type: protein_or_peptide / ID: 26 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria) / Strain: mc(2)155
Molecular weightTheoretical: 8.312485 KDa
SequenceString:
MKTGIHPEYV DTTVQCGCGH SFTTRSTKQS GTIVVEVCSQ CHPFYTGKQK ILDSGGRVAR FEKRYGKRNK AAADK

UniProtKB: Large ribosomal subunit protein bL31

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Macromolecule #27: 23S rRNA

MacromoleculeName: 23S rRNA / type: rna / ID: 27 / Number of copies: 1
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 1.01183475 MDa
SequenceString: AAGUGUUUAA GGGCGCAUGG UGGAUGCCUU GGCACUGGGA GCCGAUGAAG GACGUAGGAG GCUGCGAUAA GCCUCGGGGA GCUGUCAAC CGAGCGUUGA UCCGAGGAUG UCCGAAUGGG GAAACCCGGC ACGAGUGAUG UCGUGUCACC AGGCGCUGAA U AUAUAGGC ...String:
AAGUGUUUAA GGGCGCAUGG UGGAUGCCUU GGCACUGGGA GCCGAUGAAG GACGUAGGAG GCUGCGAUAA GCCUCGGGGA GCUGUCAAC CGAGCGUUGA UCCGAGGAUG UCCGAAUGGG GAAACCCGGC ACGAGUGAUG UCGUGUCACC AGGCGCUGAA U AUAUAGGC GUCUGGGGGG AACGCGGGGA AGUGAAACAU CUCAGUACCC GUAGGAAGAG AAAACAAAAU GUGAUUCCGU GA GUAGUGG CGAGCGAAAG CGGAGGAUGG CUAAACCGUA UGCAUGUGAU ACCGGGUAGG GGUUGUGUGU GCGGGGUUGU GGG ACCUAU CUUUCCGGCU CUACCUGGCU GGAGGGCAGU GAGAAAAUGU UGUGGUUAGC GGAAAUGGCU UGGGAUGGCC UGCC GUAGA CGGUGAGAGC CCGGUACGUG AAAACCCGAC GUCUGUCUUG AUGGUGUUCC CGAGUAGCAG CGGGCCCGUG GAAUC UGCU GUGAAUCUGC CGGGACCACC CGGUAAGCCU GAAUACUUCC CAGUGACCGA UAGCGGAUUA GUACCGUGAG GGAAUG GUG AAAAGUACCC CGGGAGGGGA GUGAAAGAGU ACCUGAAACC GUGCGCUUAC AAUCCGUCAG AGCCCUCGAC GUGUCGU GG GGUGAUGGCG UGCCUUUUGA AGAAUGAGCC UGCGAGUCAG GGACAUGUCG CGAGGUUAAC CCGGGUGGGG UAGCCGCA G CGAAAGCGAG UCUGAAUAGG GCGUAUCCAC ACAAGAGUGU GUGGUGUAGU GGUGUGUUCU GGACCCGAAG CGGAGUGAU CUACCCAUGG CCAGGGUGAA GCGCGGGUAA GACCGCGUGG AGGCCCGAAC CCACUUAGGU UGAAGACUGA GGGGAUGAGC UGUGGGUAG GGGUGAAAGG CCAAUCAAAC UCCGUGAUAG CUGGUUCUCC CCGAAAUGCA UUUAGGUGCA GCGUCGCAUG U UUCUUGCC GGAGGUAGAG CUACUGGAUG GCCGAUGGGC CCCACAGGGU UACUGACGUC AGCCAAACUC CGAAUGCCGG UA AGUCCAA GAGUGCGGCA GUGAGACGGC GGGGGAUAAG CUCCGUGCGU CGAGAGGGAA ACAGCCCAGA UCGCCGGCUA AGG CCCCUA AGCGUGUGCU AAGUGGAAAA GGAUGUGCAG UCGCGAAGAC AACCAGGAGG UUGGCUUAGA AGCAGCCACC CUUG AAAGA GUGCGUAAUA GCUCACUGGU CAAGUGAUUG UGCGCCGAUA AUGUAGCGGG GCUCAAGCAC ACCGCCGAAG CCGCG GCAG CCAACGUGUU GGCUGGGUAG GGGAGCGUCC UGCAUCCGGU GAAGCCGCCG AGUGAUCGAG UGGUGGAGGG UGUGGG AGU GAGAAUGCAG GCAUGAGUAG CGAUUAGGCA AGUGAGAACC UUGCCCGCCG AAAGACCAAG GGUUCCUGGG CCAGGCC AG UCCGCCCAGG GUGAGUCGGG ACCUAAGGCG AGGCCGACAG GCGUAGUCGA UGGACAACGG GUUGAUAUUC CCGUACCC G UGUAUGUGCG UCCAUGAUGA AUCAGCGGUA CUAACCAUCC AAAACCACCG UGACCGCACC UUUCGGGGUG UGGCGUUGG UGGGGCUGCA UGGGACCUUC GUUGGUAGUA GUCAAGCGAU GGGGUGACGC AGGAAGGUAG CCGUACCGGU CAGUGGUAAU ACCGGGGUA AGCCUGUAGG GAGUCAGAUA GGUAAAUCCG UCUGGCAUAU AUCCUGAGAG GUGAUGCAUA GCCGAGUGAG G CGAAUUCG GUGAUCCUAU GCUGCCGAGA AAAGCCUCUA GCGAGGACAU ACACGGCCCG UACCCCAAAC CAACACAGGU GG UCAGGUA GAGAAUACUA AGGCGUACGA GUGAACUAUG GUUAAGGAAC UCGGCAAAAU GCCCCCGUAA CUUCGGGAGA AGG GGGACC CACAUGGCGU GUAAGCCUUU ACGGCCCAAG CGUGAGUGGG UGGCACAAAC CAGUGAGAAG CGACUGUUUA CUAA AAACA CAGGUCCGUG CGAAGUCGCA AGACGAUGUA UACGGACUGA CGCCUGCCCG GUGCUGGAAG GUUAAGAGGA CCCGU UAAC UCCCUUUGGG GGUGAAGCGG AGAAUUUAAG CCCCAGUAAA CGGCGGUGGU AACUAUAACC AUCCUAAGGU AGCGAA AUU CCUUGUCGGG UAAGUUCCGA CCUGCACGAA UGGCGUAACG ACUUCUCAAC UGUCUCAACC AUAGACUCGG CGAAAUU GC ACUACGAGUA AAGAUGCUCG UUACGCGCGG CAGGACGAAA AGACCCCGGG ACCUUCACUA CAACUUGGUA UUGGUGCU C GAUACGGUUU GUGUAGGAUA GGUGGGAGAC UGUGAAGCUC ACACGCCAGU GUGGGUGGAG UCGUUGUUGA AAUACCACU CUGAUCGUAU UGGGCCUCUA ACCUCGGACC GUAUAUCCGG UUCAGGGACA GUGCCUGGUG GGUAGUUUAA CUGGGGCGGU UGCCUCCUA AAAUGUAACG GAGGCGCCCA AAGGUUCCCU CAACCUGGAC GGCAAUCAGG UGUUGAGUGU AAGUGCACAA G GGAGCUUG ACUGCGAGAC GGACAUGUCG AGCAGGGACG AAAGUCGGGA CUAGUGAUCC GGCACCUCUG AGUGGAAGGG GU GUCGCUC AACGGAUAAA AGGUACCCCG GGGAUAACAG GCUGAUCUUC CCCAAGAGUC CAUAUCGACG GGAUGGUUUG GCA CCUCGA UGUCGGCUCG UCGCAUCCUG GGGCUGGAGC AGGUCCCAAG GGUUGGGCUG UUCGCCCAUU AAAGCGGCAC GCGA GCUGG GUUUAGAACG UCGUGAGACA GUUCGGUCUC UAUCCGCCGC GCGCGUCAGA AGCUUGAGGA AACCUGUCCC UAGUA CGAG AGGACCGGGA CGGACGAACC UCUGGUAUAC CAGUUGUCCC ACCAGGGGCA CGGCUGGAUA GCCACGUUCG GACAGG AUA ACCGCUGAAA GCAUCUAAGC GGGAAACCUC UUCCAAGACC AGGCUUCUCA CCCUCUAGGA GGGAUAAGGC CCCCCGC AG ACCACGGGAU UGAUAGACCA GACCUGGAAG CCUAGUAAUA GGUGCAGGGA ACUGGCACUA ACCGGCCGAA AACUUAC

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Macromolecule #28: 5S rRNA

MacromoleculeName: 5S rRNA / type: rna / ID: 28 / Number of copies: 1
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria) / Strain: mc(2)155
Molecular weightTheoretical: 38.061816 KDa
SequenceString:
GUUACGGCGG UCCAUAGCGG CAGGGAAACG CCCGGUCCCA UCCCGAACCC GGAAGCUAAG CCUGCCAGCG CCGAUGAUAC UACCCAUCC GGGUGGAAAA GUAGGACACC GCCGAACAC

GENBANK: GENBANK: CP000480.1

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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

Concentration1 mg/mL
BufferpH: 7.4 / Details: 20mM HEPES, 20mM MgCl2, 100mM NH4Cl, 3mM DTT
GridModel: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 300 / Support film - Material: CARBON / Support film - topology: CONTINUOUS / Pretreatment - Type: GLOW DISCHARGE
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Instrument: FEI VITROBOT MARK IV

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: OTHER / Imaging mode: BRIGHT FIELDBright-field microscopy / Nominal defocus max: 3.0 µm / Nominal defocus min: 1.8 µm
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
Image recordingFilm or detector model: FEI FALCON III (4k x 4k) / Detector mode: INTEGRATING / Number grids imaged: 3 / Number real images: 12343 / Average exposure time: 2.0 sec. / Average electron dose: 1.34 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Particle selectionNumber selected: 1202461
Startup modelType of model: EMDB MAP
EMDB ID:

8932

Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 3.1.4)
Final 3D classificationSoftware - Name: RELION (ver. 3.1.4)
Final angle assignmentType: ANGULAR RECONSTITUTION
Final reconstructionApplied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 2.7 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 3.1.4) / Number images used: 110934
FSC plot (resolution estimation)

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Atomic model buiding 1

Initial modelPDB ID:

8whx


Chain - Source name: PDB / Chain - Initial model type: experimental model
Detailsphenix.real_space_refinement
RefinementSpace: REAL / Protocol: FLEXIBLE FIT
Output model

PDB-8why:
Cryo- EM structure of Mycobacterium smegmatis 50S ribosomal subunit (body 1) of 70S ribosome and RafH.

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