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- EMDB-27028: Cutibacterium acnes 30S ribosomal subunit with Sarecycline bound,... -

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Basic information

Entry
Database: EMDB / ID: EMD-27028
TitleCutibacterium acnes 30S ribosomal subunit with Sarecycline bound, body domain only in the local refined map
Map dataCutibacterium acnes 30S ribosomal subunit with Sarecycline bound, body domain only in the local refined map
Sample
  • Complex: 30S subunit body domain from 70S ribosome with mRNA, P-site tRNA and Sarecycline bound
    • RNA: x 2 types
    • Protein or peptide: x 13 types
  • Ligand: x 2 types
Function / homology
Function and homology information


small ribosomal subunit / tRNA binding / rRNA binding / ribosome / structural constituent of ribosome / translation / ribonucleoprotein complex / cytoplasm
Similarity search - Function
Mitochondrial mRNA-processing protein COX24, C-terminal / Mitochondrial mRNA-processing protein COX24, C-terminal / Mitochondrial domain of unknown function (DUF1713) / Ribosomal protein S16, conserved site / Ribosomal protein S16 signature. / Ribosomal protein S11, bacterial-type / Ribosomal protein S20 / Ribosomal protein S20 superfamily / Ribosomal protein S20 / Ribosomal protein S4, bacterial-type ...Mitochondrial mRNA-processing protein COX24, C-terminal / Mitochondrial mRNA-processing protein COX24, C-terminal / Mitochondrial domain of unknown function (DUF1713) / Ribosomal protein S16, conserved site / Ribosomal protein S16 signature. / Ribosomal protein S11, bacterial-type / Ribosomal protein S20 / Ribosomal protein S20 superfamily / Ribosomal protein S20 / Ribosomal protein S4, bacterial-type / 30S ribosomal protein S17 / Ribosomal protein S5, bacterial-type / Ribosomal protein S6, plastid/chloroplast / Ribosomal protein S2, bacteria/mitochondria/plastid / Ribosomal protein S18, conserved site / Ribosomal protein S18 signature. / Ribosomal protein S16 / Ribosomal protein S16 / Ribosomal protein S16 domain superfamily / Ribosomal protein S15, bacterial-type / Ribosomal protein S6 / Ribosomal protein S6 / Ribosomal protein S6 superfamily / Ribosomal protein S12, bacterial-type / Translation elongation factor EF1B/ribosomal protein S6 / Ribosomal protein S18 / Ribosomal protein S18 / Ribosomal protein S18 superfamily / Ribosomal protein S17, conserved site / Ribosomal protein S2 / Ribosomal protein S2, flavodoxin-like domain superfamily / Ribosomal protein S2 / Ribosomal protein S4/S9 N-terminal domain / Ribosomal protein S4/S9, N-terminal / Ribosomal protein S4, conserved site / Ribosomal protein S4/S9 N-terminal domain / Ribosomal protein S4/S9 / Ribosomal protein S8 / Ribosomal protein S8 superfamily / Ribosomal protein S5, N-terminal, conserved site / Ribosomal protein S5 signature. / Ribosomal protein S5 / Ribosomal protein S5, N-terminal / S5 double stranded RNA-binding domain profile. / Ribosomal protein S5, C-terminal / Ribosomal protein S5, N-terminal domain / Ribosomal protein S8 / Ribosomal protein S17 signature. / Ribosomal protein S5, C-terminal domain / S4 RNA-binding domain / RNA-binding S4 domain / RNA-binding S4 domain superfamily / Ribosomal protein S8 signature. / Ribosomal S11, conserved site / S4 domain / Ribosomal protein S17/S11 / Ribosomal protein S4 signature. / Ribosomal_S15 / Ribosomal protein S15 signature. / Ribosomal protein S15 / Ribosomal protein S12/S23 / Ribosomal protein S11 / Ribosomal protein S11 / S4 RNA-binding domain profile. / Ribosomal protein S12/S23 / Ribosomal protein S17 / Ribosomal protein S15 / Ribosomal protein S11 signature. / Ribosomal protein S12 signature. / Ribosomal protein S11 superfamily / S15/NS1, RNA-binding / Ribosomal protein S5 domain 2-type fold, subgroup / Ribosomal protein S5 domain 2-type fold / Nucleic acid-binding, OB-fold
Similarity search - Domain/homology
30S ribosomal protein S20 / Small ribosomal subunit protein uS5 / 30S ribosomal protein S11 / 30S ribosomal protein S15 / 30S ribosomal protein S2 / 30S ribosomal protein S8 / 30S ribosomal protein S16 / 30S ribosomal protein S17 / 30S ribosomal protein S4 / 30S ribosomal protein S6 ...30S ribosomal protein S20 / Small ribosomal subunit protein uS5 / 30S ribosomal protein S11 / 30S ribosomal protein S15 / 30S ribosomal protein S2 / 30S ribosomal protein S8 / 30S ribosomal protein S16 / 30S ribosomal protein S17 / 30S ribosomal protein S4 / 30S ribosomal protein S6 / 30S ribosomal protein S18 / 30S ribosomal protein S12 / AURKAIP1/COX24 domain-containing protein
Similarity search - Component
Biological speciesCutibacterium acnes (bacteria) / Escherichia coli (E. coli)
Methodsingle particle reconstruction / cryo EM / Resolution: 2.84 Å
AuthorsLomakin IB / Devarkar SC / Bunick CG
Funding support United States, 1 items
OrganizationGrant numberCountry
Almirall, LLC United States
Citation
Journal: Nucleic Acids Res / Year: 2023
Title: Sarecycline inhibits protein translation in Cutibacterium acnes 70S ribosome using a two-site mechanism.
Authors: Ivan B Lomakin / Swapnil C Devarkar / Shivali Patel / Ayman Grada / Christopher G Bunick /
Abstract: Acne vulgaris is a chronic disfiguring skin disease affecting ∼1 billion people worldwide, often having persistent negative effects on physical and mental health. The Gram-positive anaerobe, ...Acne vulgaris is a chronic disfiguring skin disease affecting ∼1 billion people worldwide, often having persistent negative effects on physical and mental health. The Gram-positive anaerobe, Cutibacterium acnes is implicated in acne pathogenesis and is, therefore, a main target for antibiotic-based acne therapy. We determined a 2.8-Å resolution structure of the 70S ribosome of Cutibacterium acnes by cryogenic electron microscopy and discovered that sarecycline, a narrow-spectrum antibiotic against Cutibacterium acnes, may inhibit two active sites of this bacterium's ribosome in contrast to the one site detected previously on the model ribosome of Thermus thermophilus. Apart from the canonical binding site at the mRNA decoding center, the second binding site for sarecycline exists at the nascent peptide exit tunnel, reminiscent of the macrolides class of antibiotics. The structure also revealed Cutibacterium acnes-specific features of the ribosomal RNA and proteins. Unlike the ribosome of the Gram-negative bacterium Escherichia coli, Cutibacterium acnes ribosome has two additional proteins, bS22 and bL37, which are also present in the ribosomes of Mycobacterium smegmatis and Mycobacterium tuberculosis. We show that bS22 and bL37 have antimicrobial properties and may be involved in maintaining the healthy homeostasis of the human skin microbiome.
#1: Journal: To be published
Title: The structure of the Cutibacterium acnes 70S ribosome
Authors: Lomakin IB / Devarkar SC / Grada A / Bunick CG
History
DepositionMay 19, 2022-
Header (metadata) releaseMar 15, 2023-
Map releaseMar 15, 2023-
UpdateApr 19, 2023-
Current statusApr 19, 2023Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_27028.png

Downloads & links

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Map

FileDownload / File: emd_27028.map.gz / Format: CCP4 / Size: 149.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationCutibacterium acnes 30S ribosomal subunit with Sarecycline bound, body domain only in the local refined map
Voxel sizeX=Y=Z: 1.068 Å
Density
Contour LevelBy AUTHOR: 0.16
Minimum - Maximum-0.48456722 - 1.3131591
Average (Standard dev.)0.0010100472 (±0.042305227)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions340340340
Spacing340340340
CellA=B=C: 363.12 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: Cutibacterium acnes 30S ribosomal subunit with Sarecycline bound,...

Fileemd_27028_half_map_1.map
AnnotationCutibacterium acnes 30S ribosomal subunit with Sarecycline bound, body domain only in the local refined map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: Cutibacterium acnes 30S ribosomal subunit with Sarecycline bound,...

Fileemd_27028_half_map_2.map
AnnotationCutibacterium acnes 30S ribosomal subunit with Sarecycline bound, body domain only in the local refined map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Sample components

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Entire : 30S subunit body domain from 70S ribosome with mRNA, P-site tRNA ...

EntireName: 30S subunit body domain from 70S ribosome with mRNA, P-site tRNA and Sarecycline bound
Components
  • Complex: 30S subunit body domain from 70S ribosome with mRNA, P-site tRNA and Sarecycline bound
    • RNA: 16S ribosomal RNA
    • Protein or peptide: 30S ribosomal protein S4
    • Protein or peptide: 30S ribosomal protein S5
    • Protein or peptide: 30S ribosomal protein S6
    • Protein or peptide: 30S ribosomal protein S8
    • Protein or peptide: 30S ribosomal protein S11
    • Protein or peptide: 30S ribosomal protein S12
    • Protein or peptide: 30S ribosomal protein S15
    • Protein or peptide: 30S ribosomal protein S17
    • Protein or peptide: 30S ribosomal protein S18
    • Protein or peptide: 30S ribosomal protein S20
    • Protein or peptide: 30S ribosomal protein S16
    • Protein or peptide: 30S ribosomal protein bS22
    • Protein or peptide: 30S ribosomal protein S2
    • RNA: mRNAMessenger RNA
  • Ligand: MAGNESIUM ION
  • Ligand: water

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Supramolecule #1: 30S subunit body domain from 70S ribosome with mRNA, P-site tRNA ...

SupramoleculeName: 30S subunit body domain from 70S ribosome with mRNA, P-site tRNA and Sarecycline bound
type: complex / ID: 1 / Chimera: Yes / Parent: 0 / Macromolecule list: #1-#15
Source (natural)Organism: Cutibacterium acnes (bacteria)

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Macromolecule #1: 16S ribosomal RNA

MacromoleculeName: 16S ribosomal RNA / type: rna / ID: 1 / Number of copies: 1
Source (natural)Organism: Cutibacterium acnes (bacteria)
Molecular weightTheoretical: 498.742875 KDa
SequenceString: UUUUUCAUUG GAGAGUUUGA UCCUGGCUCA GGACGAACGC UGGCGGCGUG CUUAACACAU GCAAGUCGAA CGGAAAGGCC CUGCUUUUG UGGGGUGCUC GAGUGGCGAA CGGGUGAGUA ACACGUGAGU AACCUGCCCU UGACUUUGGG AUAACUUCAG G AAACUGGG ...String:
UUUUUCAUUG GAGAGUUUGA UCCUGGCUCA GGACGAACGC UGGCGGCGUG CUUAACACAU GCAAGUCGAA CGGAAAGGCC CUGCUUUUG UGGGGUGCUC GAGUGGCGAA CGGGUGAGUA ACACGUGAGU AACCUGCCCU UGACUUUGGG AUAACUUCAG G AAACUGGG GCUAAUACCG GAUAGGAGCU CCUGCUGCAU GGUGGGGGUU GGAAAGUUUC GGCGGUUGGG GAUGGACUCG CG GCUUAUC AGCUUGUUGG UGGGGUAGUG GCUUACCAAG GCUUUGACGG GUAGCCGGCC UGAGAGGGUG ACCGGCCACA UUG GGACUG AGAUACGGCC CAGACUCCUA CGGGAGGCAG CAGUGGGGAA UAUUGCACAA UGGGCGGAAG CCUGAUGCAG CAAC GCCGC GUGCGGGAUG ACGGCCUUCG GGUUGUAAAC CGCUUUCGCC UGUGACGAAG CGUGAGUGAC GGUAAUGGGU AAAGA AGCA CCGGCUAACU ACG(PSU)GCCAGC AGCC(G7M)CGGUG AUACGUAGGG UGCGAGCGUU GUCCGGAUUU AUUGGGCG U AAAGGGCUCG UAGGUGGUUG AUCGCGUCGG AAGUGUAAUC UUGGGGCUUA ACCCUGAGCG UGCUUUCGAU ACGGGUUGA CUUGAGGAAG GUAGGGGAGA AUGGAAUUCC UGGUGGAGCG GUGGAAUGCG CAGAUAUCAG GAGGAACACC AGUGGCGAAG GCGGUUCUC UGGGCCUUUC CUGACGCUGA GGAGCGAAAG CGUGGGGAGC GAACAGGCUU AGAUACCCUG GUAGUCCACG C UGUAAACG GUGGGUACUA GGUGUGGGGU CCAUUCCACG GGUUCCGUGC CGUAGCUAAC GCUUUAAGUA CCCCGCCUGG GG AGUACGG CCGCAAGGCU AAAACUCAAA GGAAUUGACG GGGCCCCGCA CAAGCGGCGG AGCAUGCGGA UUAAUUCGAU (2MG)(5MC)AACGCGU AGAACCUUAC CUGGGUUUGA CAUGGAUCGG GAGUGCUCAG AGAUGGGUGU GCCUCUUUUG GGG UCGGUU CACAGGUGGU GCAUGGCUGU CGUCAGCUCG UGUCGUGAGA UGUUGGGUUA AGUCCCGCAA CGAGCGCAAC CCUU GUUCA CUGUUGCCAG CACGUUAUGG UGGGGACUCA GUGGAGACCG CCGGGGUCAA CUCGGAGGAA GGUGGGGAUG ACGUC AAGU CAUCAUGCCC CUUAUGUCCA GGGCUUCACG CAUGCUACAA UGGCUGGUAC AGAGAGUGGC GAGCCUGUGA GGGUGA GCG AAUCUCGGAA AGCCGGUCUC AGUUCGGAUU GGGGUCUGCA ACUCGACCUC AUGAAGUCGG AGUCGCUAGU AAUCGCA GA UCAGCAACGC UGCGGUGAAU ACGUUCCCGG GGCUUGUACA CAC(5MC)G(4OC)C(5MC)GU (5MC)AAGUCAUGA AA GUUGGUA ACACCCGAAG CCGGUGGCCU AACCGUUGUG GGGGAGCCGU CGAAGGUGGG ACUGGUGAUU AGGACUAAGU CG (UR3)AACAAG GUAGCCGUAC C(2MG)G(MA6)(MA6)GGUGC GGCUGGAUCA CCUCCUUUCU AAGGAG

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Macromolecule #15: mRNA

MacromoleculeName: mRNA / type: rna / ID: 15 / Number of copies: 1
Source (natural)Organism: Escherichia coli (E. coli)
Molecular weightTheoretical: 7.115308 KDa
SequenceString:
CAAGGAGGUA AAAAUGUUUU AA

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Macromolecule #2: 30S ribosomal protein S4

MacromoleculeName: 30S ribosomal protein S4 / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Cutibacterium acnes (bacteria)
Molecular weightTheoretical: 23.271605 KDa
SequenceString: MARYTGPLTK KSRRLGTDLV GNDKSFERRP YPPGVHGRGR TKDSEYSLQL REKQKARYAY GVLEKQFRRY YEEADRAQGK TGDVLLQIL ESRLDNVVYR AGLAATRRQA RQMVSHGHFL VNGKKVNIPS YRVSTHDIID VREKSKDLPP IVIARETFET R DVPAWLEV ...String:
MARYTGPLTK KSRRLGTDLV GNDKSFERRP YPPGVHGRGR TKDSEYSLQL REKQKARYAY GVLEKQFRRY YEEADRAQGK TGDVLLQIL ESRLDNVVYR AGLAATRRQA RQMVSHGHFL VNGKKVNIPS YRVSTHDIID VREKSKDLPP IVIARETFET R DVPAWLEV RPNKGRILVH QLPTRDQIVI DVNEQAIVEL YSK

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Macromolecule #3: 30S ribosomal protein S5

MacromoleculeName: 30S ribosomal protein S5 / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Cutibacterium acnes (bacteria)
Molecular weightTheoretical: 22.603695 KDa
SequenceString: MSGTQRRGGG AGGERRGRDN RRGQNDRNRN QNEYLERVVA INRVAKVVQG GRRFSFTALV VVGDGEGSVG VGYGKAKEVP AAIAKAVEE AKKHFFKVPL VGRTITHPVI GEKAAGVVML RPASPGTGVI AGGSARAVLE CAGVHDVLAK SLGSSNAINV V HATVDALQ ...String:
MSGTQRRGGG AGGERRGRDN RRGQNDRNRN QNEYLERVVA INRVAKVVQG GRRFSFTALV VVGDGEGSVG VGYGKAKEVP AAIAKAVEE AKKHFFKVPL VGRTITHPVI GEKAAGVVML RPASPGTGVI AGGSARAVLE CAGVHDVLAK SLGSSNAINV V HATVDALQ QLEEPEEVAR RRGKSVEDIA PAAMLRARKE ADEAAAAARM EEKAGVN

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Macromolecule #4: 30S ribosomal protein S6

MacromoleculeName: 30S ribosomal protein S6 / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Cutibacterium acnes (bacteria)
Molecular weightTheoretical: 11.270147 KDa
SequenceString:
MRKYEVMIII DPTVEERQVD SLMEKYLKVI TDEKGTVDNV DVWGKRRLAY DIQKKSEGIY VVVNATCEPA TIQELDRLLA IDEKIMRTK VMRPEIH

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Macromolecule #5: 30S ribosomal protein S8

MacromoleculeName: 30S ribosomal protein S8 / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Cutibacterium acnes (bacteria)
Molecular weightTheoretical: 14.67793 KDa
SequenceString:
MTMTDPIADM LTRLRNANQA YHDQTSMPHS KIKAGIAGIL KSEGYIADYK VNEPKEGEVG KTLTLTLKYG ENRERSIAGV RRISKPGLR VYAKSTALPK VLGGLGIAII STSQGLLTDK QAHEKSVGGE VLAYVW

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Macromolecule #6: 30S ribosomal protein S11

MacromoleculeName: 30S ribosomal protein S11 / type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Cutibacterium acnes (bacteria)
Molecular weightTheoretical: 14.194463 KDa
SequenceString:
MATAGHKGAP KTKVRRKEKK NVVAGQAHIK STFNNTIIAI TDPSGAVISW ASAGTVGFKG SRKSTPFAAQ MAAEAAGRRA MEHGMKRVD VFVKGPGSGR ETAIRSLGAV GLEIGPISDV TPVPHNGCRP PKRRRV

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Macromolecule #7: 30S ribosomal protein S12

MacromoleculeName: 30S ribosomal protein S12 / type: protein_or_peptide / ID: 7 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Cutibacterium acnes (bacteria)
Molecular weightTheoretical: 13.666084 KDa
SequenceString:
MPTIQQLVRK GRTDKISKNK TPALKGSPQR RGVCTRVYTT TPKKPNSALR KVARVRLSSG IEVTAYIPGV GHNLQEHSMV LVRGGRVKD LPGVRYKIVR GSLDTQGVKG RKQARSRYGA KKEK

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Macromolecule #8: 30S ribosomal protein S15

MacromoleculeName: 30S ribosomal protein S15 / type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Cutibacterium acnes (bacteria)
Molecular weightTheoretical: 10.10276 KDa
SequenceString:
MDAAEKKKII EEYATHPGDT GSPDVQVAIL TKRIAELTEH LKVHKGDHHS RRGLMLMVGQ RRRLLNYIAK NDIEHYRELI ARLGLRR

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Macromolecule #9: 30S ribosomal protein S17

MacromoleculeName: 30S ribosomal protein S17 / type: protein_or_peptide / ID: 9 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Cutibacterium acnes (bacteria)
Molecular weightTheoretical: 10.475277 KDa
SequenceString:
MSENTAERTT RRKVREGLVV SDKMNKTITV MVEDRVKHPL YGKVMTKSVR LKAHDENNEA GMGDRVRIME TRPLSATKRW RLVEIIEKA K

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Macromolecule #10: 30S ribosomal protein S18

MacromoleculeName: 30S ribosomal protein S18 / type: protein_or_peptide / ID: 10 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Cutibacterium acnes (bacteria)
Molecular weightTheoretical: 8.817497 KDa
SequenceString:
MAGPQRKSVN KKKVVPIKTV HIGAVDYKDT ALLRKFISER GKIRARRVTG LSVQDQRKVA IAIKNARELA LLPYASTAR

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Macromolecule #11: 30S ribosomal protein S20

MacromoleculeName: 30S ribosomal protein S20 / type: protein_or_peptide / ID: 11 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Cutibacterium acnes (bacteria)
Molecular weightTheoretical: 9.724277 KDa
SequenceString:
MPNIKSQIKR VKTNEKSRQR NKAVKSALRT YVRNFRRAAE AGDVEAATKA ARVANRQLDK AASKGVIHKN QAANRKSAIS KKLNSLAA

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Macromolecule #12: 30S ribosomal protein S16

MacromoleculeName: 30S ribosomal protein S16 / type: protein_or_peptide / ID: 12 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Cutibacterium acnes (bacteria)
Molecular weightTheoretical: 15.980861 KDa
SequenceString:
MATKIRLKRL GKIRTPHYRV VVMDSRAKRD GRAIEEIGQY HPKADPSVIV IDSERVQYWL GVGAQPTEAV VALLKRTGDW QKFTGDTSP SGVKPQPERP NKDDLFNAAL AEADEAPREA ITKKSEGAAA DEASESAAGD NSGEKAEA

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Macromolecule #13: 30S ribosomal protein bS22

MacromoleculeName: 30S ribosomal protein bS22 / type: protein_or_peptide / ID: 13 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Cutibacterium acnes (bacteria)
Molecular weightTheoretical: 4.120248 KDa
SequenceString:
MGSVIKKRRK RMAKKKHRKL LKRTRIQRRR AGK

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Macromolecule #14: 30S ribosomal protein S2

MacromoleculeName: 30S ribosomal protein S2 / type: protein_or_peptide / ID: 14 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Cutibacterium acnes (bacteria)
Molecular weightTheoretical: 31.583166 KDa
SequenceString: MAVVTTRQLL ESGVHFGHQT RRWNPKMKRF IFTERNGIYI IDLHQSLTYI DKAYAFVKET VAKGGQILFV GTKKQAQESI VEQATRVGM PYVNQRWLGG MLTNFQTISK RIARLKELEA MDFDKVSGSG LTKKELLMLS REKDKLEKDL GGIRDMPKVP Q AVWVVDTK ...String:
MAVVTTRQLL ESGVHFGHQT RRWNPKMKRF IFTERNGIYI IDLHQSLTYI DKAYAFVKET VAKGGQILFV GTKKQAQESI VEQATRVGM PYVNQRWLGG MLTNFQTISK RIARLKELEA MDFDKVSGSG LTKKELLMLS REKDKLEKDL GGIRDMPKVP Q AVWVVDTK KEHLAIDEAR KLKIPVVAIL DTNCDPDEVD YAIPGNDDAI RSVSLLTRII ADAAAEGLMA RSAGKSGEQP AE AEPMPDW ERELLEGDGA KTEAKAEEPK AEAKKADEAP EAEKSN

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Macromolecule #16: MAGNESIUM ION

MacromoleculeName: MAGNESIUM ION / type: ligand / ID: 16 / Number of copies: 66 / Formula: MG
Molecular weightTheoretical: 24.305 Da

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Macromolecule #17: water

MacromoleculeName: water / type: ligand / ID: 17 / Number of copies: 1 / Formula: HOH
Molecular weightTheoretical: 18.015 Da
Chemical component information

ChemComp-HOH:
WATER / Water

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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: 7.4
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 opticsC2 aperture diameter: 50.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Cs: 2.7 mm / Nominal defocus max: 2.0 µm / Nominal defocus min: 0.5 µm
Sample stageCooling holder cryogen: NITROGEN
Image recordingFilm or detector model: GATAN K3 (6k x 4k) / Average exposure time: 2.1 sec. / Average electron dose: 30.56 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Particle selectionNumber selected: 1108852
Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: cryoSPARC (ver. 3.3.1) / Software - details: Ab-initio
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: cryoSPARC (ver. 3.3.1)
Final reconstructionResolution.type: BY AUTHOR / Resolution: 2.84 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: cryoSPARC (ver. 3.3.1) / Software - details: Local Refinement / Number images used: 70853
FSC plot (resolution estimation)

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Atomic model buiding 1

RefinementSpace: REAL / Protocol: FLEXIBLE FIT
Output model

PDB-8cwo:
Cutibacterium acnes 30S ribosomal subunit with Sarecycline bound, body domain only in the local refined map

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