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- EMDB-43333: Structure of Mycobacterium smegmatis 50S ribosomal subunit bound ... -

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Basic information

Entry
Database: EMDB / ID: EMD-43333
TitleStructure of Mycobacterium smegmatis 50S ribosomal subunit bound to delNTE-HflX
Map dataStructure of Mycobacterium smegmatis 50S ribosomal subunit bound to delNTE-HflX
Sample
  • Complex: Mycobacterium smegmatis 50S ribosome bound to delNTE-HflX-GMPPCP complex
    • Protein or peptide: x 32 types
    • RNA: x 2 types
Keywordsribosome splitting / Mycobacterium smegmatis 50S / HflX / TRANSLATION / RIBOSOME
Function / homology
Function and homology information


large ribosomal subunit / transferase activity / ribosome binding / 5S rRNA binding / ribosomal large subunit assembly / large ribosomal subunit rRNA binding / cytosolic large ribosomal subunit / cytoplasmic translation / tRNA binding / negative regulation of translation ...large ribosomal subunit / transferase activity / ribosome binding / 5S rRNA binding / ribosomal large subunit assembly / large ribosomal subunit rRNA binding / cytosolic large ribosomal subunit / cytoplasmic translation / tRNA binding / negative regulation of translation / rRNA binding / ribosome / structural constituent of ribosome / translation / ribonucleoprotein complex / GTPase activity / mRNA binding / GTP binding / metal ion binding / cytoplasm
Similarity search - Function
GTPase HflX / GTPase HflX, N-terminal / HflX-type guanine nucleotide-binding (G) domain / GTP-binding protein, middle domain / GTPase HflX, N-terminal domain superfamily / GTP-binding GTPase N-terminal / GTP-binding GTPase Middle Region / HflX-type guanine nucleotide-binding (G) domain profile. / 50S ribosome-binding GTPase / GTP binding domain ...GTPase HflX / GTPase HflX, N-terminal / HflX-type guanine nucleotide-binding (G) domain / GTP-binding protein, middle domain / GTPase HflX, N-terminal domain superfamily / GTP-binding GTPase N-terminal / GTP-binding GTPase Middle Region / HflX-type guanine nucleotide-binding (G) domain profile. / 50S ribosome-binding GTPase / GTP binding domain / Ribosomal protein L25, long-form / Ribosomal protein L25, beta domain / Ribosomal protein L25, C-terminal / Ribosomal protein TL5, C-terminal domain / : / Ribosomal protein L11, bacterial-type / Ribosomal protein L31 type A / Ribosomal protein L31 signature. / Ribosomal protein L31 / Ribosomal protein L31 superfamily / Ribosomal protein L31 / Ribosomal protein L11, conserved site / Ribosomal protein L11 signature. / : / Ribosomal protein L16 signature 1. / Ribosomal protein L21, conserved site / Ribosomal protein L21 signature. / Ribosomal protein L16 signature 2. / Ribosomal protein L16, conserved site / Ribosomal protein L6, conserved site / Ribosomal protein L6 signature 1. / : / 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 L11, N-terminal / Ribosomal protein L11, N-terminal domain / Ribosomal protein L17 signature. / Ribosomal protein L11/L12 / Ribosomal protein L11, C-terminal / Ribosomal protein L11, C-terminal domain superfamily / Ribosomal protein L11/L12, N-terminal domain superfamily / Ribosomal protein L11/L12 / Ribosomal protein L11, RNA binding domain / Ribosomal L25p family / Ribosomal protein L25 / Ribosomal protein L36 signature. / Ribosomal protein L28/L24 superfamily / Ribosomal protein L25/Gln-tRNA synthetase, N-terminal / Ribosomal protein L25/Gln-tRNA synthetase, anti-codon-binding domain superfamily / Ribosomal protein L9, N-terminal domain superfamily / Ribosomal protein L9 / Ribosomal protein L9, N-terminal / Ribosomal protein L9, N-terminal domain / Ribosomal protein L33, conserved site / Ribosomal protein L33 signature. / Ribosomal protein L35, conserved site / Ribosomal protein L35 signature. / Ribosomal protein L28 / Ribosomal protein L35, non-mitochondrial / : / Ribosomal protein L5, bacterial-type / Ribosomal protein L18, bacterial-type / : / Ribosomal protein L6, bacterial-type / Ribosomal protein L9/RNase H1, N-terminal / Ribosomal protein L36 / Ribosomal protein L36 superfamily / Ribosomal protein L36 / Ribosomal protein L19, conserved site / Ribosomal protein L19 signature. / Ribosomal protein L27, conserved site / Ribosomal protein L27 signature. / Ribosomal protein L20 signature. / Ribosomal protein L22, bacterial/chloroplast-type / Ribosomal protein L14P, bacterial-type / Ribosomal protein L34, conserved site / Ribosomal protein L34 signature. / Ribosomal protein L2, bacterial/organellar-type / Ribosomal protein L35 / Ribosomal protein L35 superfamily / Ribosomal protein L35 / Ribosomal protein L33 / Ribosomal protein L33 / Ribosomal L28 family / Ribosomal protein L33 superfamily / Ribosomal protein L16 / Ribosomal protein L28/L24 / Ribosomal protein L18 / Ribosomal L18 of archaea, bacteria, mitoch. and chloroplast / Ribosomal protein L30, bacterial-type / : / L28p-like / Ribosomal protein L27 / Ribosomal L27 protein / Ribosomal protein L20 / Ribosomal L32p protein family / Ribosomal protein L19
Similarity search - Domain/homology
Large ribosomal subunit protein bL33A / Large ribosomal subunit protein uL11 / 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 uL16 / Large ribosomal subunit protein uL29 / Large ribosomal subunit protein uL14 ...Large ribosomal subunit protein bL33A / Large ribosomal subunit protein uL11 / 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 uL16 / 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 uL30 / Large ribosomal subunit protein uL15 / Large ribosomal subunit protein bL36 / Large ribosomal subunit protein bL17 / Large ribosomal subunit protein uL13 / Uncharacterized protein / Large ribosomal subunit protein bL19 / GTPase HflX / 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 bL25 / Large ribosomal subunit protein bL32 / Large ribosomal subunit protein bL9 / Large ribosomal subunit protein bL34 / Large ribosomal subunit protein uL18 / Large ribosomal subunit protein bL28
Similarity search - Component
Biological speciesMycolicibacterium smegmatis MC2 155 (bacteria)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.44 Å
AuthorsMajumdar S / Koripella RK / Sharma MR / Manjari SR / Banavali NK / Agrawal RK
Funding support United States, 3 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)R01 GM61576 United States
National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)R01Al1554732 United States
National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)R01AI132422 United States
CitationJournal: Proc Natl Acad Sci U S A / Year: 2025
Title: HflX-mediated drug resistance through ribosome splitting and rRNA disordering in mycobacteria.
Authors: Soneya Majumdar / Amuliya Kashyap / Ravi K Koripella / Manjuli R Sharma / Kelley Hurst-Hess / Swati R Manjari / Nilesh K Banavali / Pallavi Ghosh / Rajendra K Agrawal /
Abstract: HflX is a highly conserved ribosome-associated GTPase implicated in rescuing stalled ribosomes and mediating antibiotic resistance in several bacteria, including macrolide-lincosamide antibiotic ...HflX is a highly conserved ribosome-associated GTPase implicated in rescuing stalled ribosomes and mediating antibiotic resistance in several bacteria, including macrolide-lincosamide antibiotic resistance in mycobacteria. Mycobacterial HflXs carry a distinct N-terminal extension (NTE) and a small insertion, as compared to their eubacterial homologs. Here, we present several high-resolution cryo-EM structures of mycobacterial HflX in complex with the 70S ribosome and its 50S subunit, with and without antibiotics. These structures reveal a distinct mechanism for HflX-mediated ribosome splitting and antibiotic resistance in mycobacteria. Our findings indicate that the NTE of mycobacterial HflX induces persistent disordering of multiple 23S rRNA helices, facilitating the dissociation of the 70S ribosome and generating an inactive pool of 50S subunits. During this process, HflX undergoes a large conformational change that stabilizes its NTE. Mycobacterial HflX also acts as an anti-association factor by binding to predissociated 50S subunits. Our structures show that a mycobacteria-specific insertion in HflX reaches far into the peptidyl transferase center (PTC), such that it would overlap with the ribosome-bound macrolide antibiotics. However, in the presence of antibiotics, this insertion retracts, adjusts around, and interacts with the antibiotic molecules. These results suggest that mycobacterial HflX is agnostic to antibiotic presence in the PTC. It mediates antibiotic resistance by splitting antibiotic-stalled 70S ribosomes and inactivating the resulting 50S subunits.
History
DepositionJan 10, 2024-
Header (metadata) releaseFeb 19, 2025-
Map releaseFeb 19, 2025-
UpdateApr 30, 2025-
Current statusApr 30, 2025Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_43333.png

Downloads & links

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Map

FileDownload / File: emd_43333.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationStructure of Mycobacterium smegmatis 50S ribosomal subunit bound to delNTE-HflX
Projections & slices

Image control

Size
Brightness
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AxesZ (Sec.)Y (Row.)X (Col.)
0.85 Å/pix.
x 400 pix.
= 338.4 Å		0.85 Å/pix.
x 400 pix.
= 338.4 Å		0.85 Å/pix.
x 400 pix.
= 338.4 Å

Surface

Projections

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Images are generated by Spider.

Voxel sizeX=Y=Z: 0.846 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.25
Minimum - Maximum-0.33529183 - 1.4421017
Average (Standard dev.)0.029989785 (±0.09088484)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions400400400
Spacing400400400
CellA=B=C: 338.4 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: Structure of Mycobacterium smegmatis 50S ribosomal subunit bound...

Fileemd_43333_half_map_1.map
AnnotationStructure of Mycobacterium smegmatis 50S ribosomal subunit bound to delNTE-HflX half map A
Projections & Slices
AxesZYX

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Histogram (log scale)

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Half map: Structure of Mycobacterium smegmatis 50S ribosomal subunit bound...

Fileemd_43333_half_map_2.map
AnnotationStructure of Mycobacterium smegmatis 50S ribosomal subunit bound to delNTE-HflX half map B
Projections & Slices
AxesZYX

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Histogram

Histogram (log scale)

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Sample components

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Entire : Mycobacterium smegmatis 50S ribosome bound to delNTE-HflX-GMPPCP ...

EntireName: Mycobacterium smegmatis 50S ribosome bound to delNTE-HflX-GMPPCP complex
Components
  • Complex: Mycobacterium smegmatis 50S ribosome bound to delNTE-HflX-GMPPCP complex
    • Protein or peptide: 50S ribosomal protein L30
    • Protein or peptide: 50S Ribosomal Protein L37
    • Protein or peptide: GTPase HflX
    • RNA: 23S ribosomal RNA
    • RNA: 5S ribosomal RNA
    • 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 L11
    • Protein or peptide: 50S Ribosomal Protein L13
    • Protein or peptide: 50S ribosomal protein L14
    • Protein or peptide: 50S ribosomal protein L15
    • Protein or peptide: Large ribosomal subunit protein uL16
    • 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 L25
    • 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 L32
    • Protein or peptide: 50S Ribosomal Protein L33
    • Protein or peptide: 50S ribosomal protein L34
    • Protein or peptide: 50S ribosomal protein L35
    • Protein or peptide: 50S ribosomal protein L36
    • Protein or peptide: 50S Ribosomal Protein L31

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Supramolecule #1: Mycobacterium smegmatis 50S ribosome bound to delNTE-HflX-GMPPCP ...

SupramoleculeName: Mycobacterium smegmatis 50S ribosome bound to delNTE-HflX-GMPPCP complex
type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#34
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)

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Macromolecule #1: 50S ribosomal protein L30

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

UniProtKB: Large ribosomal subunit protein uL30

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Macromolecule #2: 50S Ribosomal Protein L37

MacromoleculeName: 50S Ribosomal Protein L37 / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 2.841375 KDa
SequenceString:
MAKRGRKKRD RKHSKANHGK RPNA

UniProtKB: Uncharacterized protein

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Macromolecule #3: GTPase HflX

MacromoleculeName: GTPase HflX / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 46.516578 KDa
Recombinant expressionOrganism: Escherichia coli 'BL21-Gold(DE3)pLysS AG' (bacteria)
SequenceString: EYRQLRLERV VLVGVWTEGS AADAEASLAE LAALAETAGS EVLEGLIQRR DKPDPSTYIG SGKAAELREV VLATGADTVI CDGELSPAQ LNALEKAVKV KVIDRTALIL DIFAQHATSR EGKAQVSLAQ MEYMLPRLRG WGESMSRQAG GRAGGAGGGV G TRGPGETK ...String:
EYRQLRLERV VLVGVWTEGS AADAEASLAE LAALAETAGS EVLEGLIQRR DKPDPSTYIG SGKAAELREV VLATGADTVI CDGELSPAQ LNALEKAVKV KVIDRTALIL DIFAQHATSR EGKAQVSLAQ MEYMLPRLRG WGESMSRQAG GRAGGAGGGV G TRGPGETK IETDRRRIRE RMAKLRREIR DMKKIRDTQR GSRRRSEIPS VAIVGYTNAG KSSLLNALTG AGVLVENALF AT LEPTTRR GEFEDGRPFV LTDTVGFVRH LPTQLVEAFR STLEEVVDAD LLIHVVDGSD VNPLAQINAV RTVINEVVAE YDI APPPEL LVVNKIDAAT GVGLAQLRRA LPDAVFVSAR TGDGLDKLRS RMGELVESTD ATVDVTIPYD RGDLVARVHT DGHV DATEH TDAGTRIKAR VPAPLAATLR EYATFA

UniProtKB: GTPase HflX

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Macromolecule #6: 50S ribosomal protein L2

MacromoleculeName: 50S ribosomal protein L2 / type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
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 #7: 50S ribosomal protein L3

MacromoleculeName: 50S ribosomal protein L3 / type: protein_or_peptide / ID: 7 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
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 #8: 50S Ribosomal Protein L4

MacromoleculeName: 50S Ribosomal Protein L4 / type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
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 #9: 50S Ribosomal Protein L5

MacromoleculeName: 50S Ribosomal Protein L5 / type: protein_or_peptide / ID: 9 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
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 #10: 50S ribosomal protein L6

MacromoleculeName: 50S ribosomal protein L6 / type: protein_or_peptide / ID: 10 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
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 #11: 50S ribosomal protein L9

MacromoleculeName: 50S ribosomal protein L9 / type: protein_or_peptide / ID: 11 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
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 #12: 50S ribosomal protein L11

MacromoleculeName: 50S ribosomal protein L11 / type: protein_or_peptide / ID: 12 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 15.023374 KDa
SequenceString:
MAPKKKVAGL IKLQIQAGQA NPAPPVGPAL GQHGVNIMEF CKAYNAATES QRGNVIPVEI TVYEDRSFTF ALKTPPAAKL LLKAAGVQK GSGEPHKTKV AKVTWDQVRE IAETKKADLN ANDIDAAAKI IAGTARSMGI TVE

UniProtKB: Large ribosomal subunit protein uL11

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Macromolecule #13: 50S Ribosomal Protein L13

MacromoleculeName: 50S Ribosomal Protein L13 / type: protein_or_peptide / ID: 13 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
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 #14: 50S ribosomal protein L14

MacromoleculeName: 50S ribosomal protein L14 / type: protein_or_peptide / ID: 14 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
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 #15: 50S ribosomal protein L15

MacromoleculeName: 50S ribosomal protein L15 / type: protein_or_peptide / ID: 15 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
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 #16: Large ribosomal subunit protein uL16

MacromoleculeName: Large ribosomal subunit protein uL16 / type: protein_or_peptide / ID: 16 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 15.77424 KDa
SequenceString:
MLIPRKVKHR KQHHPEQRGI ASGGTSVSFG DYGIQALEHA YITNRQIESA RIAINRHIKR GGKVWINIFP DRPLTKKPAE TRMGSGKGS PEWWVANVKP GRVLFELSYP DEKTARDALT RAIHKLPIKA RIVTREEQF

UniProtKB: Large ribosomal subunit protein uL16

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Macromolecule #17: 50S ribosomal protein L17

MacromoleculeName: 50S ribosomal protein L17 / type: protein_or_peptide / ID: 17 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
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 #18: 50S Ribosomal Protein L18

MacromoleculeName: 50S Ribosomal Protein L18 / type: protein_or_peptide / ID: 18 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
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 #19: 50S ribosomal protein L19

MacromoleculeName: 50S ribosomal protein L19 / type: protein_or_peptide / ID: 19 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
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 #20: 50S Ribosomal Protein L20

MacromoleculeName: 50S Ribosomal Protein L20 / type: protein_or_peptide / ID: 20 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
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 #21: 50S Ribosomal Protein L21

MacromoleculeName: 50S Ribosomal Protein L21 / type: protein_or_peptide / ID: 21 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
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 #22: 50S Ribosomal Protein L22

MacromoleculeName: 50S Ribosomal Protein L22 / type: protein_or_peptide / ID: 22 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
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 #23: 50S Ribosomal Protein L23

MacromoleculeName: 50S Ribosomal Protein L23 / type: protein_or_peptide / ID: 23 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
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 #24: 50S ribosomal protein L24

MacromoleculeName: 50S ribosomal protein L24 / type: protein_or_peptide / ID: 24 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
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 #25: 50S ribosomal protein L25

MacromoleculeName: 50S ribosomal protein L25 / type: protein_or_peptide / ID: 25 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 22.571078 KDa
SequenceString: MAKTANIPNK LTANVRTRTG KGASRQARRD GKVPAVLYGH GTDPQHLELN ARDFAAVLRS HGTNAILTLD IEGTEQLALT KALDVHPIR RNIQHADLLV VQRGEKVTVE VTVLVEGDAT PGTLVTQDAN TIEIEAEALS IPEQLTVSVE GVEAGTQITA G QISLPEGV ...String:
MAKTANIPNK LTANVRTRTG KGASRQARRD GKVPAVLYGH GTDPQHLELN ARDFAAVLRS HGTNAILTLD IEGTEQLALT KALDVHPIR RNIQHADLLV VQRGEKVTVE VTVLVEGDAT PGTLVTQDAN TIEIEAEALS IPEQLTVSVE GVEAGTQITA G QISLPEGV NLISDPELLV VNVVEAPSAE ALEEEGAGET GAAEAPAEEA PAEEPAE

UniProtKB: Large ribosomal subunit protein bL25

+
Macromolecule #26: 50S ribosomal protein L27

MacromoleculeName: 50S ribosomal protein L27 / type: protein_or_peptide / ID: 26 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 9.249582 KDa
SequenceString:
MAHKKGASSS RNGRDSAAQR LGVKRFGGQV VKAGEILVRQ RGTHFHPGVN VGRGGDDTLF ALAPGAVEFG AKRGRKTVNI VPVARPEA

UniProtKB: Large ribosomal subunit protein bL27

+
Macromolecule #27: 50S Ribosomal Protein L28

MacromoleculeName: 50S Ribosomal Protein L28 / type: protein_or_peptide / ID: 27 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 6.891016 KDa
SequenceString:
MAAVCDICGK GPGFGKSVSH SHRRTSRRWN PNIQPVRAVT RPGGNKQRIN ACTSCIKAGK VSRA

UniProtKB: Large ribosomal subunit protein bL28

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Macromolecule #28: 50S ribosomal protein L29

MacromoleculeName: 50S ribosomal protein L29 / type: protein_or_peptide / ID: 28 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
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 #29: 50S ribosomal protein L32

MacromoleculeName: 50S ribosomal protein L32 / type: protein_or_peptide / ID: 29 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 6.467719 KDa
SequenceString:
MAVPKRRMSR ANTRSRRAQW KAEAPGLVTV SVAGQQRKVP RRLLKAARLG LVDLDKR

UniProtKB: Large ribosomal subunit protein bL32

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Macromolecule #30: 50S Ribosomal Protein L33

MacromoleculeName: 50S Ribosomal Protein L33 / type: protein_or_peptide / ID: 30 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 6.468501 KDa
SequenceString:
MASSTDVRPK ITLACEVCKH RNYITKKNRR NDPDRLEIKK FCPNCGTHQP HKESR

UniProtKB: Large ribosomal subunit protein bL33A

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Macromolecule #31: 50S ribosomal protein L34

MacromoleculeName: 50S ribosomal protein L34 / type: protein_or_peptide / ID: 31 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 5.52258 KDa
SequenceString:
MAKGKRTFQP NNRRRARVHG FRLRMRTRAG RAIVANRRSK GRRALTA

UniProtKB: Large ribosomal subunit protein bL34

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Macromolecule #32: 50S ribosomal protein L35

MacromoleculeName: 50S ribosomal protein L35 / type: protein_or_peptide / ID: 32 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 7.298505 KDa
SequenceString:
MPKAKTHSGA SKRFRRTGTG KIVRQKANRR HLLEHKPTKR TRRLDGRTTV SAADNSRINK LLNG

UniProtKB: Large ribosomal subunit protein bL35

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Macromolecule #33: 50S ribosomal protein L36

MacromoleculeName: 50S ribosomal protein L36 / type: protein_or_peptide / ID: 33 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 4.341277 KDa
SequenceString:
MKVNPSVKPI CDKCRVIRRH GRVMVICSDP RHKQRQG

UniProtKB: Large ribosomal subunit protein bL36

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Macromolecule #34: 50S Ribosomal Protein L31

MacromoleculeName: 50S Ribosomal Protein L31 / type: protein_or_peptide / ID: 34 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
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 #4: 23S ribosomal RNA

MacromoleculeName: 23S ribosomal RNA / type: rna / ID: 4 / Number of copies: 1
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
Molecular weightTheoretical: 1.012140938 MDa
SequenceString: UAAGUGUUUA AGGGCGCAUG GUGGAUGCCU UGGCACUGGG AGCCGAUGAA GGACGUAGGA GGCUGCGAUA AGCCUCGGGG AGCUGUCAA CCGAGCGUUG AUCCGAGGAU GUCCGAAUGG GGAAACCCGG CACGAGUGAU GUCGUGUCAC CAGGCGCUGA A UAUAUAGG ...String:
UAAGUGUUUA AGGGCGCAUG GUGGAUGCCU UGGCACUGGG AGCCGAUGAA GGACGUAGGA GGCUGCGAUA AGCCUCGGGG AGCUGUCAA CCGAGCGUUG AUCCGAGGAU GUCCGAAUGG GGAAACCCGG CACGAGUGAU GUCGUGUCAC CAGGCGCUGA A UAUAUAGG CGUCUGGGGG GAACGCGGGG AAGUGAAACA UCUCAGUACC CGUAGGAAGA GAAAACAAAA UGUGAUUCCG UG AGUAGUG GCGAGCGAAA GCGGAGGAUG GCUAAACCGU AUGCAUGUGA UACCGGGUAG GGGUUGUGUG UGCGGGGUUG UGG GACCUA UCUUUCCGGC UCUACCUGGC UGGAGGGCAG UGAGAAAAUG UUGUGGUUAG CGGAAAUGGC UUGGGAUGGC CUGC CGUAG ACGGUGAGAG CCCGGUACGU GAAAACCCGA CGUCUGUCUU GAUGGUGUUC CCGAGUAGCA GCGGGCCCGU GGAAU CUGC UGUGAAUCUG CCGGGACCAC CCGGUAAGCC UGAAUACUUC CCAGUGACCG AUAGCGGAUU AGUACCGUGA GGGAAU GGU GAAAAGUACC CCGGGAGGGG AGUGAAAGAG UACCUGAAAC CGUGCGCUUA CAAUCCGUCA GAGCCCUCGA CGUGUCG UG GGGUGAUGGC GUGCCUUUUG AAGAAUGAGC CUGCGAGUCA GGGACAUGUC GCGAGGUUAA CCCGGGUGGG GUAGCCGC A GCGAAAGCGA GUCUGAAUAG GGCGUAUCCA CACAAGAGUG UGUGGUGUAG UGGUGUGUUC UGGACCCGAA GCGGAGUGA UCUACCCAUG GCCAGGGUGA AGCGCGGGUA AGACCGCGUG GAGGCCCGAA CCCACUUAGG UUGAAGACUG AGGGGAUGAG CUGUGGGUA GGGGUGAAAG GCCAAUCAAA CUCCGUGAUA GCUGGUUCUC CCCGAAAUGC AUUUAGGUGC AGCGUCGCAU G UUUCUUGC CGGAGGUAGA GCUACUGGAU GGCCGAUGGG CCCCACAGGG UUACUGACGU CAGCCAAACU CCGAAUGCCG GU AAGUCCA AGAGUGCGGC AGUGAGACGG CGGGGGAUAA GCUCCGUGCG UCGAGAGGGA AACAGCCCAG AUCGCCGGCU AAG GCCCCU AAGCGUGUGC UAAGUGGAAA AGGAUGUGCA GUCGCGAAGA CAACCAGGAG GUUGGCUUAG AAGCAGCCAC CCUU GAAAG AGUGCGUAAU AGCUCACUGG UCAAGUGAUU GUGCGCCGAU AAUGUAGCGG GGCUCAAGCA CACCGCCGAA GCCGC GGCA GCCAACGUGU UGGCUGGGUA GGGGAGCGUC CUGCAUCCGG UGAAGCCGCC GAGUGAUCGA GUGGUGGAGG GUGUGG GAG UGAGAAUGCA GGCAUGAGUA GCGAUUAGGC AAGUGAGAAC CUUGCCCGCC GAAAGACCAA GGGUUCCUGG GCCAGGC CA GUCCGCCCAG GGUGAGUCGG GACCUAAGGC GAGGCCGACA GGCGUAGUCG AUGGACAACG GGUUGAUAUU CCCGUACC C GUGUAUGUGC GUCCAUGAUG AAUCAGCGGU ACUAACCAUC CAAAACCACC GUGACCGCAC CUUUCGGGGU GUGGCGUUG GUGGGGCUGC AUGGGACCUU CGUUGGUAGU AGUCAAGCGA UGGGGUGACG CAGGAAGGUA GCCGUACCGG UCAGUGGUAA UACCGGGGU AAGCCUGUAG GGAGUCAGAU AGGUAAAUCC GUCUGGCAUA UAUCCUGAGA GGUGAUGCAU AGCCGAGUGA G GCGAAUUC GGUGAUCCUA UGCUGCCGAG AAAAGCCUCU AGCGAGGACA UACACGGCCC GUACCCCAAA CCAACACAGG UG GUCAGGU AGAGAAUACU AAGGCGUACG AGUGAACUAU GGUUAAGGAA CUCGGCAAAA UGCCCCCGUA ACUUCGGGAG AAG GGGGAC CCACAUGGCG UGUAAGCCUU UACGGCCCAA GCGUGAGUGG GUGGCACAAA CCAGUGAGAA GCGACUGUUU ACUA AAAAC ACAGGUCCGU GCGAAGUCGC AAGACGAUGU AUACGGACUG ACGCCUGCCC GGUGCUGGAA GGUUAAGAGG ACCCG UUAA CUCCCUUUGG GGGUGAAGCG GAGAAUUUAA GCCCCAGUAA ACGGCGGUGG UAACUAUAAC CAUCCUAAGG UAGCGA AAU UCCUUGUCGG GUAAGUUCCG ACCUGCACGA AUGGCGUAAC GACUUCUCAA CUGUCUCAAC CAUAGACUCG GCGAAAU UG CACUACGAGU AAAGAUGCUC GUUACGCGCG GCAGGACGAA AAGACCCCGG GACCUUCACU ACAACUUGGU AUUGGUGC U CGAUACGGUU UGUGUAGGAU AGGUGGGAGA CUGUGAAGCU CACACGCCAG UGUGGGUGGA GUCGUUGUUG AAAUACCAC UCUGAUCGUA UUGGGCCUCU AACCUCGGAC CGUAUAUCCG GUUCAGGGAC AGUGCCUGGU GGGUAGUUUA ACUGGGGCGG UUGCCUCCU AAAAUGUAAC GGAGGCGCCC AAAGGUUCCC UCAACCUGGA CGGCAAUCAG GUGUUGAGUG UAAGUGCACA A GGGAGCUU GACUGCGAGA CGGACAUGUC GAGCAGGGAC GAAAGUCGGG ACUAGUGAUC CGGCACCUCU GAGUGGAAGG GG UGUCGCU CAACGGAUAA AAGGUACCCC GGGGAUAACA GGCUGAUCUU CCCCAAGAGU CCAUAUCGAC GGGAUGGUUU GGC ACCUCG AUGUCGGCUC GUCGCAUCCU GGGGCUGGAG CAGGUCCCAA GGGUUGGGCU GUUCGCCCAU UAAAGCGGCA CGCG AGCUG GGUUUAGAAC GUCGUGAGAC AGUUCGGUCU CUAUCCGCCG CGCGCGUCAG AAGCUUGAGG AAACCUGUCC CUAGU ACGA GAGGACCGGG ACGGACGAAC CUCUGGUAUA CCAGUUGUCC CACCAGGGGC ACGGCUGGAU AGCCACGUUC GGACAG GAU AACCGCUGAA AGCAUCUAAG CGGGAAACCU CUUCCAAGAC CAGGCUUCUC ACCCUCUAGG AGGGAUAAGG CCCCCCG CA GACCACGGGA UUGAUAGACC AGACCUGGAA GCCUAGUAAU AGGUGCAGGG AACUGGCACU AACCGGCCGA AAACUUAC

GENBANK: GENBANK: CP000480.1

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Macromolecule #5: 5S ribosomal RNA

MacromoleculeName: 5S ribosomal RNA / type: rna / ID: 5 / Number of copies: 1
Source (natural)Organism: Mycolicibacterium smegmatis MC2 155 (bacteria)
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

BufferpH: 7.5
Component:
ConcentrationFormulaName
20.0 mMC8H18N2O4SHEPES
30.0 mMNH4ClAmmonium chloride
10.0 mMMgCl2Magnesium chloride
5.0 mMC2H6OSbeta-mercaptoethanol

Details: 20 mM HEPES-K at pH 7.5, 30 mM ammonium chloride, 10 mM magnesium chloride, and 5 mM beta-mercaptoethanol
GridModel: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 300 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 30 sec. / Pretreatment - Atmosphere: AIR
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277.15 K / Instrument: FEI VITROBOT MARK IV

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Image recordingFilm or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Digitization - Frames/image: 1-28 / Number grids imaged: 1 / Number real images: 8700 / Average electron dose: 60.64 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 0.001 mm / Nominal defocus max: 2.5 µm / Nominal defocus min: 0.8 µm / Nominal magnification: 81000
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Particle selectionNumber selected: 332311
Startup modelType of model: NONE
Final reconstructionApplied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 3.44 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 27550
Initial angle assignmentType: NOT APPLICABLE
Final angle assignmentType: PROJECTION MATCHING
FSC plot (resolution estimation)

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Atomic model buiding 1

Initial model
PDB IDChain

6dzi

source_name: PDB, initial_model_type: experimental model

5o61

source_name: PDB, initial_model_type: experimental model
RefinementSpace: REAL / Protocol: FLEXIBLE FIT / Overall B value: 67.4 / Target criteria: Cross-correlation coefficient
Output model

PDB-8vkw:
Structure of Mycobacterium smegmatis 50S ribosomal subunit bound to delNTE-HflX

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