EMDB-12635: Initiated 70S ribosome in complex with 2A protein from encephalom...

Basic information

• Entry: Database: EMDB / ID: EMD-12635
• Title: Initiated 70S ribosome in complex with 2A protein from encephalomyocarditis virus (EMCV)
• Map data: Post-processed, masked map weighted by local resolution

Sample

• Complex: Initiated 70S ribosome in complex with 2A protein from encephalomyocarditis virus (EMCV)
  • Protein or peptide: x 52 types
  • RNA: x 4 types
• RNA: x 1 types
• Ligand: x 4 types

• Keywords: 70S ribosome / initiation complex / EMCV / cardiovirus / 2A / picornavirus / frameshifting / PRF / RNA-binding protein / protein-mediated frameshifting / ribosome-binding protein / beta-shell / viral protein / RIBOSOME

Function / homology

Function:

positive stranded viral RNA replication / host cell nucleolus / stringent response / symbiont-mediated suppression of host cytoplasmic pattern recognition receptor signaling pathway via inhibition of RIG-I activity / misfolded RNA binding / Group I intron splicing / RNA folding / transcriptional attenuation / endoribonuclease inhibitor activity / RNA-binding transcription regulator activity / positive regulation of ribosome biogenesis / negative regulation of cytoplasmic translation / translational termination / DnaA-L2 complex / translation repressor activity / negative regulation of DNA-templated DNA replication initiation / mRNA regulatory element binding translation repressor activity / ribosome assembly / assembly of large subunit precursor of preribosome / picornain 3C / positive regulation of RNA splicing / cytosolic ribosome assembly / T=pseudo3 icosahedral viral capsid / response to reactive oxygen species / regulation of cell growth / DNA-templated transcription termination / host cell cytoplasmic vesicle membrane / maintenance of translational fidelity / response to radiation / mRNA 5'-UTR binding / symbiont-mediated suppression of host gene expression / ribosomal small subunit biogenesis / small ribosomal subunit rRNA binding / large ribosomal subunit / channel activity / ribosome biogenesis / ribosome binding / ribosomal small subunit assembly / small ribosomal subunit / 5S rRNA binding / large ribosomal subunit rRNA binding / transferase activity / cytosolic small ribosomal subunit / monoatomic ion transmembrane transport / ribosomal large subunit assembly / cytoplasmic translation / cytosolic large ribosomal subunit / tRNA binding / RNA helicase activity / negative regulation of translation / rRNA binding / RNA helicase / ribosome / structural constituent of ribosome / induction by virus of host autophagy / symbiont entry into host cell / ribonucleoprotein complex / translation / RNA-directed RNA polymerase / viral translational frameshifting / cysteine-type endopeptidase activity / response to antibiotic / RNA-dependent RNA polymerase activity / negative regulation of DNA-templated transcription / mRNA binding / virus-mediated perturbation of host defense response / DNA-templated transcription / virion attachment to host cell / structural molecule activity / ATP hydrolysis activity / proteolysis / DNA binding / RNA binding / zinc ion binding / ATP binding / membrane / metal ion binding / cytosol / cytoplasm

Domain/homology:

Leader peptide, picornavirus / Viral leader polypeptide zinc finger / Virion protein N terminal domain / Capsid protein VP4, Picornavirus / Viral protein VP4 subunit / Capsid protein VP4 superfamily, Picornavirus / Helicase/polymerase/peptidase polyprotein, Calicivirus-type / Ribosomal protein L10, eubacterial, conserved site / Ribosomal protein L10 signature. / Ribosomal protein L10 / Picornavirus coat protein / Ribosomal protein S21, conserved site / Ribosomal protein S21 signature. / Ribosomal protein L25, short-form / Ribosomal protein S14, bacterial/plastid / Ribosomal protein L11, bacterial-type / Ribosomal protein L31 type A / Ribosomal protein S21 superfamily / Ribosomal protein S21 / Ribosomal protein S16, conserved site / Ribosomal protein S16 signature. / Ribosomal protein S21 / 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 L21, conserved site / Ribosomal protein L21 signature. / Ribosomal protein L10-like domain superfamily / Ribosomal protein L10P / Ribosomal protein L10 / Ribosomal protein L16 signature 1. / : / Ribosomal protein L6, conserved site / Ribosomal protein L6 signature 1. / Peptidase C3A/C3B, picornaviral / 3C cysteine protease (picornain 3C) / Picornavirales 3C/3C-like protease domain / Picornavirales 3C/3C-like protease domain profile. / Ribosomal protein L16, conserved site / Ribosomal protein L16 signature 2. / 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 L17 signature. / Picornavirus capsid / picornavirus capsid protein / Helicase, superfamily 3, single-stranded RNA virus / Superfamily 3 helicase of positive ssRNA viruses domain profile. / Ribosomal protein L11, N-terminal / Ribosomal L25p family / Ribosomal protein L11, N-terminal domain / Ribosomal protein L25 / Helicase, superfamily 3, single-stranded DNA/RNA virus / RNA helicase / 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, RNA binding domain / Ribosomal protein L11/L12 / 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 L32p, bacterial type / 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 L18, bacterial-type / Ribosomal protein L6, bacterial-type / Ribosomal protein L9/RNase H1, N-terminal / Ribosomal protein L19, conserved site / Ribosomal protein L19 signature. / Ribosomal protein S3, bacterial-type / Ribosomal protein S6, conserved site / Ribosomal protein S6 signature. / Ribosomal protein S19, bacterial-type / Picornavirus/Calicivirus coat protein / Ribosomal protein S13, bacterial-type / Ribosomal protein L36 / Ribosomal protein L36 superfamily / Ribosomal protein L36 / Ribosomal protein S7, bacterial/organellar-type / Ribosomal protein S11, bacterial-type

Component:

50S ribosomal protein L10 / Small ribosomal subunit protein bS18 / Small ribosomal subunit protein uS10 / Small ribosomal subunit protein uS19 / Small ribosomal subunit protein uS3 / Small ribosomal subunit protein uS17 / Small ribosomal subunit protein uS14 / Small ribosomal subunit protein uS8 / Small ribosomal subunit protein uS5 / Small ribosomal subunit protein uS13 / Small ribosomal subunit protein uS11 / Small ribosomal subunit protein uS4 / Small ribosomal subunit protein uS9 / Small ribosomal subunit protein uS15 / Small ribosomal subunit protein bS21 / Small ribosomal subunit protein bS16 / Small ribosomal subunit protein uS2 / Small ribosomal subunit protein bS20 / Small ribosomal subunit protein bS6 / Small ribosomal subunit protein uS7 / Large ribosomal subunit protein uL15 / Large ribosomal subunit protein uL11 / Large ribosomal subunit protein bL19 / Large ribosomal subunit protein bL20 / Large ribosomal subunit protein bL27 / Large ribosomal subunit protein bL28 / Large ribosomal subunit protein uL29 / Large ribosomal subunit protein bL32 / Large ribosomal subunit protein bL33 / Large ribosomal subunit protein bL34 / Large ribosomal subunit protein bL35 / Large ribosomal subunit protein bL36A / Large ribosomal subunit protein bL9 / Small ribosomal subunit protein uS12 / Large ribosomal subunit protein uL13 / Large ribosomal subunit protein uL14 / Large ribosomal subunit protein uL16 / Large ribosomal subunit protein uL23 / Large ribosomal subunit protein bL17 / Large ribosomal subunit protein bL21 / Large ribosomal subunit protein uL30 / Large ribosomal subunit protein uL6 / Large ribosomal subunit protein uL18 / Genome polyprotein / Large ribosomal subunit protein uL2 / Large ribosomal subunit protein uL3 / Large ribosomal subunit protein uL24 / Large ribosomal subunit protein uL4 / Large ribosomal subunit protein uL22 / Large ribosomal subunit protein uL5 / Large ribosomal subunit protein bL25 / Large ribosomal subunit protein bL31

• Biological species: Escherichia coli (E. coli) / Mengo encephalomyocarditis virus / Encephalomyocarditis virus
• Method: single particle reconstruction / cryo EM / Resolution: 2.66 Å
• Authors: Hill CH / Napthine S

Funding support

United Kingdom, European Union, 3 items

Organization	Grant number	Country
Wellcome Trust	202797/Z/16/Z	United Kingdom
Wellcome Trust	098406/Z/12/B	United Kingdom
European Communitys Seventh Framework Programme	BIOSTRUCT-X (Contract No. 283570)	European Union

• Citation: Journal: Nat Commun / Year: 2021; Title: Structural and molecular basis for Cardiovirus 2A protein as a viral gene expression switch.; Authors: Chris H Hill / Lukas Pekarek / Sawsan Napthine / Anuja Kibe / Andrew E Firth / Stephen C Graham / Neva Caliskan / Ian Brierley / ; Abstract: Programmed -1 ribosomal frameshifting (PRF) in cardioviruses is activated by the 2A protein, a multi-functional virulence factor that also inhibits cap-dependent translational initiation. Here we present the X-ray crystal structure of 2A and show that it selectively binds to a pseudoknot-like conformation of the PRF stimulatory RNA element in the viral genome. Using optical tweezers, we demonstrate that 2A stabilises this RNA element, likely explaining the increase in PRF efficiency in the presence of 2A. Next, we demonstrate a strong interaction between 2A and the small ribosomal subunit and present a cryo-EM structure of 2A bound to initiated 70S ribosomes. Multiple copies of 2A bind to the 16S rRNA where they may compete for binding with initiation and elongation factors. Together, these results define the structural basis for RNA recognition by 2A, show how 2A-mediated stabilisation of an RNA pseudoknot promotes PRF, and reveal how 2A accumulation may shut down translation during virus infection.

History

Deposition	Mar 17, 2021	-
Header (metadata) release	Dec 15, 2021	-
Map release	Dec 15, 2021	-
Update	Apr 24, 2024	-
Current status	Apr 24, 2024	Processing site: PDBe / Status: Released

Map

• File: Download / File: emd_12635.map.gz / Format: CCP4 / Size: 347.6 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
• Annotation: Post-processed, masked map weighted by local resolution
• Voxel size: X=Y=Z: 1.07 Å

Density

Contour Level	By AUTHOR: 0.05 / Movie #1: 0.05
Minimum - Maximum	-0.4982394 - 1.0113602
Average (Standard dev.)	0.00007548818 (±0.020886237)

• Symmetry: Space group: 1

Details

EMDB XML:

Map geometry	Axis order X Y Z Origin 0 0 0 Dimensions 450 450 450 Spacing 450 450 450
Cell	A=B=C: 481.50003 Å α=β=γ: 90.0 °

CCP4 map header:

mode	Image stored as Reals
Å/pix. X/Y/Z	1.07	1.07	1.07
M x/y/z	450	450	450
origin x/y/z	0.000	0.000	0.000
length x/y/z	481.500	481.500	481.500
α/β/γ	90.000	90.000	90.000
MAP C/R/S	1	2	3
start NC/NR/NS	0	0	0
NC/NR/NS	450	450	450
D min/max/mean	-0.498	1.011	0.000

Supplemental data

Mask #1

• File: emd_12635_msk_1.map

Additional map: Unmasked map from Refine3D

• File: emd_12635_additional_1.map
• Annotation: Unmasked map from Refine3D

Additional map: Unmasked, unfiltered half-map (1) from Refine3D

• File: emd_12635_additional_2.map
• Annotation: Unmasked, unfiltered half-map (1) from Refine3D

Additional map: Unmasked, unfiltered half-map (2) from Refine3D

• File: emd_12635_additional_3.map
• Annotation: Unmasked, unfiltered half-map (2) from Refine3D

Sample components

Entire : Initiated 70S ribosome in complex with 2A protein from encephalom...

• Entire: Name: Initiated 70S ribosome in complex with 2A protein from encephalomyocarditis virus (EMCV)

Components

• Complex: Initiated 70S ribosome in complex with 2A protein from encephalomyocarditis virus (EMCV)
  • 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 L10
  • 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: 50S ribosomal protein L16
  • 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 L30
  • RNA: 23S ribosomal RNA
  • RNA: 16S ribosomal RNA
  • RNA: 5S ribosomal RNA
  • Protein or peptide: 50S ribosomal protein L31
  • 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: 30S ribosomal protein S2
  • Protein or peptide: 30S ribosomal protein S3
  • 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 S7
  • Protein or peptide: 30S ribosomal protein S8
  • Protein or peptide: 30S ribosomal protein S9
  • Protein or peptide: 30S ribosomal protein S10
  • Protein or peptide: 30S ribosomal protein S11
  • Protein or peptide: 30S ribosomal protein S12
  • Protein or peptide: 30S ribosomal protein S13
  • Protein or peptide: 30S ribosomal protein S14
  • Protein or peptide: 30S ribosomal protein S15
  • Protein or peptide: 30S ribosomal protein S16
  • Protein or peptide: 30S ribosomal protein S17
  • Protein or peptide: 30S ribosomal protein S18
  • Protein or peptide: 30S ribosomal protein S19
  • Protein or peptide: 30S ribosomal protein S20
  • Protein or peptide: 30S ribosomal protein S21
  • RNA: mRNA
  • Protein or peptide: Protein 2A
• RNA: fMet-NH-tRNA(fMet)
• Ligand: MAGNESIUM ION
• Ligand: N-FORMYLMETHIONINE
• Ligand: ZINC ION
• Ligand: water

Supramolecule #1: Initiated 70S ribosome in complex with 2A protein from encephalom...

• Supramolecule: Name: Initiated 70S ribosome in complex with 2A protein from encephalomyocarditis virus (EMCV); type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#28, #30-#57; Details: 70S ribosome subunits and initiator fMet-tRNA purified from E.coli. mRNA template was generated by in vitro transcription EMCV 2A protein was recombinantly expressed in E.coli

Macromolecule #1: 50S ribosomal protein L2

• Macromolecule: Name: 50S ribosomal protein L2 / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 29.923619 KDa

Sequence

String:

MAVVKCKPTS PGRRHVVKVV NPELHKGKPF APLLEKNSKS GGRNNNGRIT TRHIGGGHKQ AYRIVDFKRN KDGIPAVVER LEYDPNRSA NIALVLYKDG ERRYILAPKG LKAGDQIQSG VDAAIKPGNT LPMRNIPVGS TVHNVEMKPG KGGQLARSAG T YVQIVARD GAYVTLRLRS GEMRKVEADC RATLGEVGNA EHMLRVLGKA GAARWRGVRP TVRGTAMNPV DHPHGGGEGR NF GKHPVTP WGVQTKGKKT RSNKRTDKFI VRRRSK

UniProtKB: Large ribosomal subunit protein uL2

Macromolecule #2: 50S ribosomal protein L3

• Macromolecule: Name: 50S ribosomal protein L3 / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 22.277535 KDa

Sequence

String:

MIGLVGKKVG MTRIFTEDGV SIPVTVIEVE ANRVTQVKDL ANDGYRAIQV TTGAKKANRV TKPEAGHFAK AGVEAGRGLW EFRLAEGEE FTVGQSISVE LFADVKKVDV TGTSKGKGFA GTVKRWNFRT QDATHGNSLS HRVPGSIGQN QTPGKVFKGK K MAGQMGNE RVTVQSLDVV RVDAERNLLL VKGAVPGATG SDLIVKPAVK A

UniProtKB: Large ribosomal subunit protein uL3

Macromolecule #3: 50S ribosomal protein L4

• Macromolecule: Name: 50S ribosomal protein L4 / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 22.121566 KDa

Sequence

String:

MELVLKDAQS ALTVSETTFG RDFNEALVHQ VVVAYAAGAR QGTRAQKTRA EVTGSGKKPW RQKGTGRARS GSIKSPIWRS GGVTFAARP QDHSQKVNKK MYRGALKSIL SELVRQDRLI VVEKFSVEAP KTKLLAQKLK DMALEDVLII TGELDENLFL A ARNLHKVD VRDATGIDPV SLIAFDKVVM TADAVKQVEE MLA

UniProtKB: Large ribosomal subunit protein uL4

Macromolecule #4: 50S ribosomal protein L5

• Macromolecule: Name: 50S ribosomal protein L5 / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 20.333611 KDa

Sequence

String:

MAKLHDYYKD EVVKKLMTEF NYNSVMQVPR VEKITLNMGV GEAIADKKLL DNAAADLAAI SGQKPLITKA RKSVAGFKIR QGYPIGCKV TLRGERMWEF FERLITIAVP RIRDFRGLSA KSFDGRGNYS MGVREQIIFP EIDYDKVDRV RGLDITITTT A KSDEEGRA LLAAFDFPFR K

UniProtKB: Large ribosomal subunit protein uL5

Macromolecule #5: 50S ribosomal protein L6

• Macromolecule: Name: 50S ribosomal protein L6 / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 18.932791 KDa

Sequence

String:

MSRVAKAPVV VPAGVDVKIN GQVITIKGKN GELTRTLNDA VEVKHADNTL TFGPRDGYAD GWAQAGTARA LLNSMVIGVT EGFTKKLQL VGVGYRAAVK GNVINLSLGF SHPVDHQLPA GITAECPTQT EIVLKGADKQ VIGQVAADLR AYRRPEPYKG K GVRYADEV VRTKEAKKK

UniProtKB: Large ribosomal subunit protein uL6

Macromolecule #6: 50S ribosomal protein L9

• Macromolecule: Name: 50S ribosomal protein L9 / type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 15.78902 KDa

Sequence

String:

MQVILLDKVA NLGSLGDQVN VKAGYARNFL VPQGKAVPAT KKNIEFFEAR RAELEAKLAE VLAAANARAE KINALETVTI ASKAGDEGK LFGSIGTRDI ADAVTAAGVE VAKSEVRLPN GVLRTTGEHE VSFQVHSEVF AKVIVNVVAE

UniProtKB: Large ribosomal subunit protein bL9

Macromolecule #7: 50S ribosomal protein L10

• Macromolecule: Name: 50S ribosomal protein L10 / type: protein_or_peptide / ID: 7 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 17.736596 KDa

Sequence

String:

MALNLQDKQA IVAEVSEVAK GALSAVVADS RGVTVDKMTE LRKAGREAGV YMRVVRNTLL RRAVEGTPFE CLKDAFVGPT LIAYSMEHP GAAARLFKEF AKANAKFEVK AAAFEGELIP ASQIDRLATL PTYEEAIARL MATMKEASAG KLVRTLAAVR D AKEAA

UniProtKB: 50S ribosomal protein L10

Macromolecule #8: 50S ribosomal protein L11

• Macromolecule: Name: 50S ribosomal protein L11 / type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 14.894362 KDa

Sequence

String:

MAKKVQAYVK LQVAAGMANP SPPVGPALGQ QGVNIMEFCK AFNAKTDSIE KGLPIPVVIT VYADRSFTFV TKTPPAAVLL KKAAGIKSG SGKPNKDKVG KISRAQLQEI AQTKAADMTG ADIEAMTRSI EGTARSMGLV VED

UniProtKB: Large ribosomal subunit protein uL11

Macromolecule #9: 50S ribosomal protein L13

• Macromolecule: Name: 50S ribosomal protein L13 / type: protein_or_peptide / ID: 9 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 16.050606 KDa

Sequence

String:

MKTFTAKPET VKRDWYVVDA TGKTLGRLAT ELARRLRGKH KAEYTPHVDT GDYIIVLNAD KVAVTGNKRT DKVYYHHTGH IGGIKQATF EEMIARRPER VIEIAVKGML PKGPLGRAMF RKLKVYAGNE HNHAAQQPQV LDI

UniProtKB: Large ribosomal subunit protein uL13

Macromolecule #10: 50S ribosomal protein L14

• Macromolecule: Name: 50S ribosomal protein L14 / type: protein_or_peptide / ID: 10 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 13.565067 KDa

Sequence

String:

MIQEQTMLNV ADNSGARRVM CIKVLGGSHR RYAGVGDIIK ITIKEAIPRG KVKKGDVLKA VVVRTKKGVR RPDGSVIRFD GNACVLLNN NSEQPIGTRI FGPVTRELRS EKFMKIISLA PEVL

UniProtKB: Large ribosomal subunit protein uL14

Macromolecule #11: 50S ribosomal protein L15

• Macromolecule: Name: 50S ribosomal protein L15 / type: protein_or_peptide / ID: 11 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 15.008471 KDa

Sequence

String:

MRLNTLSPAE GSKKAGKRLG RGIGSGLGKT GGRGHKGQKS RSGGGVRRGF EGGQMPLYRR LPKFGFTSRK AAITAEIRLS DLAKVEGGV VDLNTLKAAN IIGIQIEFAK VILAGEVTTP VTVRGLRVTK GARAAIEAAG GKIEE

UniProtKB: Large ribosomal subunit protein uL15

Macromolecule #12: 50S ribosomal protein L16

• Macromolecule: Name: 50S ribosomal protein L16 / type: protein_or_peptide / ID: 12 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 15.312269 KDa

Sequence

String:

MLQPKRTKFR KMHKGRNRGL AQGTDVSFGS FGLKAVGRGR LTARQIEAAR RAMTRAVKRQ GKIWIRVFPD KPITEKPLAV RMGKGKGNV EYWVALIQPG KVLYEMDGVP EELAREAFKL AAAKLPIKTT FVTKTVM

UniProtKB: Large ribosomal subunit protein uL16

Macromolecule #13: 50S ribosomal protein L17

• Macromolecule: Name: 50S ribosomal protein L17 / type: protein_or_peptide / ID: 13 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 14.393657 KDa

Sequence

String:

MRHRKSGRQL NRNSSHRQAM FRNMAGSLVR HEIIKTTLPK AKELRRVVEP LITLAKTDSV ANRRLAFART RDNEIVAKLF NELGPRFAS RAGGYTRILK CGFRAGDNAP MAYIELVDRS EKAEAAAE

UniProtKB: Large ribosomal subunit protein bL17

Macromolecule #14: 50S ribosomal protein L18

• Macromolecule: Name: 50S ribosomal protein L18 / type: protein_or_peptide / ID: 14 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 12.794668 KDa

Sequence

String:

MDKKSARIRR ATRARRKLQE LGATRLVVHR TPRHIYAQVI APNGSEVLVA ASTVEKAIAE QLKYTGNKDA AAAVGKAVAE RALEKGIKD VSFDRSGFQY HGRVQALADA AREAGLQF

UniProtKB: Large ribosomal subunit protein uL18

Macromolecule #15: 50S ribosomal protein L19

• Macromolecule: Name: 50S ribosomal protein L19 / type: protein_or_peptide / ID: 15 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 13.159278 KDa

Sequence

String:

MSNIIKQLEQ EQMKQDVPSF RPGDTVEVKV WVVEGSKKRL QAFEGVVIAI RNRGLHSAFT VRKISNGEGV ERVFQTHSPV VDSISVKRR GAVRKAKLYY LRERTGKAAR IKERLN

UniProtKB: Large ribosomal subunit protein bL19

Macromolecule #16: 50S ribosomal protein L20

• Macromolecule: Name: 50S ribosomal protein L20 / type: protein_or_peptide / ID: 16 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 13.528024 KDa

Sequence

String:

MARVKRGVIA RARHKKILKQ AKGYYGARSR VYRVAFQAVI KAGQYAYRDR RQRKRQFRQL WIARINAAAR QNGISYSKFI NGLKKASVE IDRKILADIA VFDKVAFTAL VEKAKAALA

UniProtKB: Large ribosomal subunit protein bL20

Macromolecule #17: 50S ribosomal protein L21

• Macromolecule: Name: 50S ribosomal protein L21 / type: protein_or_peptide / ID: 17 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 11.586374 KDa

Sequence

String:

MYAVFQSGGK QHRVSEGQTV RLEKLDIATG ETVEFAEVLM IANGEEVKIG VPFVDGGVIK AEVVAHGRGE KVKIVKFRRR KHYRKQQGH RQWFTDVKIT GISA

UniProtKB: Large ribosomal subunit protein bL21

Macromolecule #18: 50S ribosomal protein L22

• Macromolecule: Name: 50S ribosomal protein L22 / type: protein_or_peptide / ID: 18 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 12.253359 KDa

Sequence

String:

METIAKHRHA RSSAQKVRLV ADLIRGKKVS QALDILTYTN KKAAVLVKKV LESAIANAEH NDGADIDDLK VTKIFVDEGP SMKRIMPRA KGRADRILKR TSHITVVVSD R

UniProtKB: Large ribosomal subunit protein uL22

Macromolecule #19: 50S ribosomal protein L23

• Macromolecule: Name: 50S ribosomal protein L23 / type: protein_or_peptide / ID: 19 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 11.22216 KDa

Sequence

String:

MIREERLLKV LRAPHVSEKA STAMEKSNTI VLKVAKDATK AEIKAAVQKL FEVEVEVVNT LVVKGKVKRH GQRIGRRSDW KKAYVTLKE GQNLDFVGGA E

UniProtKB: Large ribosomal subunit protein uL23

Macromolecule #20: 50S ribosomal protein L24

• Macromolecule: Name: 50S ribosomal protein L24 / type: protein_or_peptide / ID: 20 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 11.33925 KDa

Sequence

String:

MAAKIRRDDE VIVLTGKDKG KRGKVKNVLS SGKVIVEGIN LVKKHQKPVP ALNQPGGIVE KEAAIQVSNV AIFNAATGKA DRVGFRFED GKKVRFFKSN SETIK

UniProtKB: Large ribosomal subunit protein uL24

Macromolecule #21: 50S ribosomal protein L25

• Macromolecule: Name: 50S ribosomal protein L25 / type: protein_or_peptide / ID: 21 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 10.713465 KDa

Sequence

String:

MFTINAEVRK EQGKGASRRL RAANKFPAII YGGKEAPLAI ELDHDKVMNM QAKAEFYSEV LTIVVDGKEI KVKAQDVQRH PYKPKLQHI DFVRA

UniProtKB: Large ribosomal subunit protein bL25

Macromolecule #22: 50S ribosomal protein L27

• Macromolecule: Name: 50S ribosomal protein L27 / type: protein_or_peptide / ID: 22 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 9.14654 KDa

Sequence

String:

MAHKKAGGST RNGRDSEAKR LGVKRFGGES VLAGSIIVRQ RGTKFHAGAN VGCGRDHTLF AKADGKVKFE VKGPKNRKFI SIEAE

UniProtKB: Large ribosomal subunit protein bL27

Macromolecule #23: 50S ribosomal protein L28

• Macromolecule: Name: 50S ribosomal protein L28 / type: protein_or_peptide / ID: 23 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 9.027551 KDa

Sequence

String:

MSRVCQVTGK RPVTGNNRSH ALNATKRRFL PNLHSHRFWV ESEKRFVTLR VSAKGMRVID KKGIDTVLAE LRARGEKY

UniProtKB: Large ribosomal subunit protein bL28

Macromolecule #24: 50S ribosomal protein L29

• Macromolecule: Name: 50S ribosomal protein L29 / type: protein_or_peptide / ID: 24 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 7.286464 KDa

Sequence

String:

MKAKELREKS VEELNTELLN LLREQFNLRM QAASGQLQQS HLLKQVRRDV ARVKTLLNEK AGA

UniProtKB: Large ribosomal subunit protein uL29

Macromolecule #25: 50S ribosomal protein L30

• Macromolecule: Name: 50S ribosomal protein L30 / type: protein_or_peptide / ID: 25 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 6.55482 KDa

Sequence

String:

MAKTIKITQT RSAIGRLPKH KATLLGLGLR RIGHTVERED TPAIRGMINA VSFMVKVEE

UniProtKB: Large ribosomal subunit protein uL30

Macromolecule #30: 50S ribosomal protein L31

• Macromolecule: Name: 50S ribosomal protein L31 / type: protein_or_peptide / ID: 30 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 7.887117 KDa

Sequence

String:

MKKDIHPKYE EITASCSCGN VMKIRSTVGH DLNLDVCSKC HPFFTGKQRD VATGGRVDRF NKRFNIPGSK

UniProtKB: Large ribosomal subunit protein bL31

Macromolecule #31: 50S ribosomal protein L32

• Macromolecule: Name: 50S ribosomal protein L32 / type: protein_or_peptide / ID: 31 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 6.463445 KDa

Sequence

String:

MAVQQNKPTR SKRGMRRSHD ALTAVTSLSV DKTSGEKHLR HHITADGYYR GRKVIAK

UniProtKB: Large ribosomal subunit protein bL32

Macromolecule #32: 50S ribosomal protein L33

• Macromolecule: Name: 50S ribosomal protein L33 / type: protein_or_peptide / ID: 32 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 6.388631 KDa

Sequence

String:

MAKGIREKIK LVSSAGTGHF YTTTKNKRTK PEKLELKKFD PVVRQHVIYK EAKIK

UniProtKB: Large ribosomal subunit protein bL33

Macromolecule #33: 50S ribosomal protein L34

• Macromolecule: Name: 50S ribosomal protein L34 / type: protein_or_peptide / ID: 33 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 5.397463 KDa

Sequence

String:

MKRTFQPSVL KRNRSHGFRA RMATKNGRQV LARRRAKGRA RLTVSK

UniProtKB: Large ribosomal subunit protein bL34

Macromolecule #34: 50S ribosomal protein L35

• Macromolecule: Name: 50S ribosomal protein L35 / type: protein_or_peptide / ID: 34 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 7.313032 KDa

Sequence

String:

MPKIKTVRGA AKRFKKTGKG GFKHKHANLR HILTKKATKR KRHLRPKAMV SKGDLGLVIA CLPYA

UniProtKB: Large ribosomal subunit protein bL35

Macromolecule #35: 50S ribosomal protein L36

• Macromolecule: Name: 50S ribosomal protein L36 / type: protein_or_peptide / ID: 35 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 4.37739 KDa

Sequence

String:

MKVRASVKKL CRNCKIVKRD GVIRVICSAE PKHKQRQG

UniProtKB: Large ribosomal subunit protein bL36A

Macromolecule #36: 30S ribosomal protein S2

• Macromolecule: Name: 30S ribosomal protein S2 / type: protein_or_peptide / ID: 36 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 26.78167 KDa

Sequence

String:

MATVSMRDML KAGVHFGHQT RYWNPKMKPF IFGARNKVHI INLEKTVPMF NEALAELNKI ASRKGKILFV GTKRAASEAV KDAALSCDQ FFVNHRWLGG MLTNWKTVRQ SIKRLKDLET QSQDGTFDKL TKKEALMRTR ELEKLENSLG GIKDMGGLPD A LFVIDADH EHIAIKEANN LGIPVFAIVD TNSDPDGVDF VIPGNDDAIR AVTLYLGAVA ATVREGRSQD LASQAEESFV EA E

UniProtKB: Small ribosomal subunit protein uS2

Macromolecule #37: 30S ribosomal protein S3

• Macromolecule: Name: 30S ribosomal protein S3 / type: protein_or_peptide / ID: 37 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 26.031316 KDa

Sequence

String:

MGQKVHPNGI RLGIVKPWNS TWFANTKEFA DNLDSDFKVR QYLTKELAKA SVSRIVIERP AKSIRVTIHT ARPGIVIGKK GEDVEKLRK VVADIAGVPA QINIAEVRKP ELDAKLVADS ITSQLERRVM FRRAMKRAVQ NAMRLGAKGI KVEVSGRLGG A EIARTEWY REGRVPLHTL RADIDYNTSE AHTTYGVIGV KVWIFKGEIL GGMAAVEQPE KPAAQPKKQQ RKGRK

UniProtKB: Small ribosomal subunit protein uS3

Macromolecule #38: 30S ribosomal protein S4

• Macromolecule: Name: 30S ribosomal protein S4 / type: protein_or_peptide / ID: 38 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 23.514199 KDa

Sequence

String:

MARYLGPKLK LSRREGTDLF LKSGVRAIDT KCKIEQAPGQ HGARKPRLSD YGVQLREKQK VRRIYGVLER QFRNYYKEAA RLKGNTGEN LLALLEGRLD NVVYRMGFGA TRAEARQLVS HKAIMVNGRV VNIASYQVSP NDVVSIREKA KKQSRVKAAL E LAEQREKP TWLEVDAGKM EGTFKRKPER SDLSADINEH LIVELYSK

UniProtKB: Small ribosomal subunit protein uS4

Macromolecule #39: 30S ribosomal protein S5

• Macromolecule: Name: 30S ribosomal protein S5 / type: protein_or_peptide / ID: 39 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 17.629398 KDa

Sequence

String:

MAHIEKQAGE LQEKLIAVNR VSKTVKGGRI FSFTALTVVG DGNGRVGFGY GKAREVPAAI QKAMEKARRN MINVALNNGT LQHPVKGVH TGSRVFMQPA SEGTGIIAGG AMRAVLEVAG VHNVLAKAYG STNPINVVRA TIDGLENMNS PEMVAAKRGK S VEEILGK

UniProtKB: Small ribosomal subunit protein uS5

Macromolecule #40: 30S ribosomal protein S6

• Macromolecule: Name: 30S ribosomal protein S6 / type: protein_or_peptide / ID: 40 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 15.727512 KDa

Sequence

String:

MRHYEIVFMV HPDQSEQVPG MIERYTAAIT GAEGKIHRLE DWGRRQLAYP INKLHKAHYV LMNVEAPQEV IDELETTFRF NDAVIRSMV MRTKHAVTEA SPMVKAKDER RERRDDFANE TADDAEAGDS EEEEEE

UniProtKB: Small ribosomal subunit protein bS6

Macromolecule #41: 30S ribosomal protein S7

• Macromolecule: Name: 30S ribosomal protein S7 / type: protein_or_peptide / ID: 41 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli) / Strain: K12
• Molecular weight: Theoretical: 20.055156 KDa

Sequence

String:

MPRRRVIGQR KILPDPKFGS ELLAKFVNIL MVDGKKSTAE SIVYSALETL AQRSGKSELE AFEVALENVR PTVEVKSRRV GGSTYQVPV EVRPVRRNAL AMRWIVEAAR KRGDKSMALR LANELSDAAE NKGTAVKKRE DVHRMAEANK AFAHYRWLSL R SFSHQAGA SSKQPALGYL N

UniProtKB: Small ribosomal subunit protein uS7

Macromolecule #42: 30S ribosomal protein S8

• Macromolecule: Name: 30S ribosomal protein S8 / type: protein_or_peptide / ID: 42 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 14.146557 KDa

Sequence

String:

MSMQDPIADM LTRIRNGQAA NKAAVTMPSS KLKVAIANVL KEEGFIEDFK VEGDTKPELE LTLKYFQGKA VVESIQRVSR PGLRIYKRK DELPKVMAGL GIAVVSTSKG VMTDRAARQA GLGGEIICYV A

UniProtKB: Small ribosomal subunit protein uS8

Macromolecule #43: 30S ribosomal protein S9

• Macromolecule: Name: 30S ribosomal protein S9 / type: protein_or_peptide / ID: 43 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 14.88627 KDa

Sequence

String:

MAENQYYGTG RRKSSAARVF IKPGNGKIVI NQRSLEQYFG RETARMVVRQ PLELVDMVEK LDLYITVKGG GISGQAGAIR HGITRALME YDESLRSELR KAGFVTRDAR QVERKKVGLR KARRRPQFSK R

UniProtKB: Small ribosomal subunit protein uS9

Macromolecule #44: 30S ribosomal protein S10

• Macromolecule: Name: 30S ribosomal protein S10 / type: protein_or_peptide / ID: 44 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 11.755597 KDa

Sequence

String:

MQNQRIRIRL KAFDHRLIDQ ATAEIVETAK RTGAQVRGPI PLPTRKERFT VLISPHVNKD ARDQYEIRTH LRLVDIVEPT EKTVDALMR LDLAAGVDVQ ISLG

UniProtKB: Small ribosomal subunit protein uS10

Macromolecule #45: 30S ribosomal protein S11

• Macromolecule: Name: 30S ribosomal protein S11 / type: protein_or_peptide / ID: 45 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 13.870975 KDa

Sequence

String:

MAKAPIRARK RVRKQVSDGV AHIHASFNNT IVTITDRQGN ALGWATAGGS GFRGSRKSTP FAAQVAAERC ADAVKEYGIK NLEVMVKGP GPGRESTIRA LNAAGFRITN ITDVTPIPHN GCRPPKKRRV

UniProtKB: Small ribosomal subunit protein uS11

Macromolecule #46: 30S ribosomal protein S12

• Macromolecule: Name: 30S ribosomal protein S12 / type: protein_or_peptide / ID: 46 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli) / Strain: K12
• Molecular weight: Theoretical: 13.814249 KDa

Sequence

String:

MATVNQLVRK PRARKVAKSN VPALEACPQK RGVCTRVYTT TPKKPNSALR KVCRVRLTNG FEVTSYIGGE GHNLQEHSVI LIRGGRVK(0TD) LPGVRYHTVR GALDCSGVKD RKQARSKYGV KRPKA

UniProtKB: Small ribosomal subunit protein uS12

Macromolecule #47: 30S ribosomal protein S13

• Macromolecule: Name: 30S ribosomal protein S13 / type: protein_or_peptide / ID: 47 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 13.128467 KDa

Sequence

String:

MARIAGINIP DHKHAVIALT SIYGVGKTRS KAILAAAGIA EDVKISELSE GQIDTLRDEV AKFVVEGDLR REISMSIKRL MDLGCYRGL RHRRGLPVRG QRTKTNARTR KGPRKPIKK

UniProtKB: Small ribosomal subunit protein uS13

Macromolecule #48: 30S ribosomal protein S14

• Macromolecule: Name: 30S ribosomal protein S14 / type: protein_or_peptide / ID: 48 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 11.60656 KDa

Sequence

String:

MAKQSMKARE VKRVALADKY FAKRAELKAI ISDVNASDED RWNAVLKLQT LPRDSSPSRQ RNRCRQTGRP HGFLRKFGLS RIKVREAAM RGEIPGLKKA SW

UniProtKB: Small ribosomal subunit protein uS14

Macromolecule #49: 30S ribosomal protein S15

• Macromolecule: Name: 30S ribosomal protein S15 / type: protein_or_peptide / ID: 49 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 10.290816 KDa

Sequence

String:

MSLSTEATAK IVSEFGRDAN DTGSTEVQVA LLTAQINHLQ GHFAEHKKDH HSRRGLLRMV SQRRKLLDYL KRKDVARYTQ LIERLGLRR

UniProtKB: Small ribosomal subunit protein uS15

Macromolecule #50: 30S ribosomal protein S16

• Macromolecule: Name: 30S ribosomal protein S16 / type: protein_or_peptide / ID: 50 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 9.207572 KDa

Sequence

String:

MVTIRLARHG AKKRPFYQVV VADSRNARNG RFIERVGFFN PIASEKEEGT RLDLDRIAHW VGQGATISDR VAALIKEVNK AA

UniProtKB: Small ribosomal subunit protein bS16

Macromolecule #51: 30S ribosomal protein S17

• Macromolecule: Name: 30S ribosomal protein S17 / type: protein_or_peptide / ID: 51 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 9.724491 KDa

Sequence

String:

MTDKIRTLQG RVVSDKMEKS IVVAIERFVK HPIYGKFIKR TTKLHVHDEN NECGIGDVVE IRECRPLSKT KSWTLVRVVE KAVL

UniProtKB: Small ribosomal subunit protein uS17

Macromolecule #52: 30S ribosomal protein S18

• Macromolecule: Name: 30S ribosomal protein S18 / type: protein_or_peptide / ID: 52 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 9.005472 KDa

Sequence

String:

MARYFRRRKF CRFTAEGVQE IDYKDIATLK NYITESGKIV PSRITGTRAK YQRQLARAIK RARYLSLLPY TDRHQ

UniProtKB: Small ribosomal subunit protein bS18

Macromolecule #53: 30S ribosomal protein S19

• Macromolecule: Name: 30S ribosomal protein S19 / type: protein_or_peptide / ID: 53 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 10.455355 KDa

Sequence

String:

MPRSLKKGPF IDLHLLKKVE KAVESGDKKP LRTWSRRSTI FPNMIGLTIA VHNGRQHVPV FVTDEMVGHK LGEFAPTRTY RGHAADKKA KKK

UniProtKB: Small ribosomal subunit protein uS19

Macromolecule #54: 30S ribosomal protein S20

• Macromolecule: Name: 30S ribosomal protein S20 / type: protein_or_peptide / ID: 54 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 9.708464 KDa

Sequence

String:

MANIKSAKKR AIQSEKARKH NASRRSMMRT FIKKVYAAIE AGDKAAAQKA FNEMQPIVDR QAAKGLIHKN KAARHKANLT AQINKLA

UniProtKB: Small ribosomal subunit protein bS20

Macromolecule #55: 30S ribosomal protein S21

• Macromolecule: Name: 30S ribosomal protein S21 / type: protein_or_peptide / ID: 55 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 8.524039 KDa

Sequence

String:

MPVIKVRENE PFDVALRRFK RSCEKAGVLA EVRRREFYEK PTTERKRAKA SAVKRHAKKL ARENARRTRL Y

UniProtKB: Small ribosomal subunit protein bS21

Macromolecule #57: Protein 2A

• Macromolecule: Name: Protein 2A / type: protein_or_peptide / ID: 57 / Number of copies: 3 / Enantiomer: LEVO
• Source (natural): Organism: Mengo encephalomyocarditis virus
• Molecular weight: Theoretical: 17.831182 KDa
• Recombinant expression: Organism: Escherichia coli (E. coli)

Sequence

String:

SPNPLDVSKT YPTLHILLQF NHRGLEARIF RHGQLWAETH AEVVLRSKTK QISFLSNGSY PSMDATTPLN PWKSTYQAVL RAEPHRVTM DVYHKRIRPF RLPLVQKEWR TCEENVFGLY HVFETHYAGY FSDLLIHDVE TNPGGSKHHH HHH

UniProtKB: Genome polyprotein

Macromolecule #26: 23S ribosomal RNA

• Macromolecule: Name: 23S ribosomal RNA / type: rna / ID: 26 / Number of copies: 1
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 941.832625 KDa

Sequence

String:

GGUUAAGCGA CUAAGCGUAC ACGGUGGAUG CCCUGGCAGU CAGAGGCGAU GAAGGACGUG CUAAUCUGCG AUAAGCGUCG GUAAGGUGA UAUGAACCGU UAUAACCGGC GAUUUCCGAA UGGGGAAACC CAGUGUGUUU CGACACACUA UCAUUAACUG A AUCCAUAG GUUAAUGAGG CGAACCGGGG GAACUGAAAC AUCUAAGUAC CCCGAGGAAA AGAAAUCAAC CGAGAUUCCC CC AGUAGCG GCGAGCGAAC GGGGAGCAGC CCAGAGCCUG AAUCAGUGUG UGUGUUAGUG GAAGCGUCUG GAAAGGCGCG CGA UACAGG GUGACAGCCC CGUACACAAA AAUGCACAUG CUGUGAGCUC GAUGAGUAGG GCGGGACACG UGGUAUCCUG UCUG AAUAU GGGGGGACCA UCCUCCAAGG CUAAAUACUC CUGACUGACC GAUAGUGAAC CAGUACCGUG AGGGAAAGGC GAAAA GAAC CCCGGCGAGG GGAGUGAAAA AGAACCUGAA ACCGUGUACG UACAAGCAGU GGGAGCACGC UUAGGCGUGU GACUGC GUA CCUUUUGUAU AAUGGGUCAG CGACUUAUAU UCUGUAGCAA GGUUAACCGA AUAGGGGAGC CGAAGGGAAA CCGAGUC UU AACUGGGCGU UAAGUUGCAG GGUAUAGACC CGAAACCCGG UGAUCUAGCC AUGGGCAGGU UGAAGGUUGG GUAACACU A ACUGGAGGAC CGAACCGACU AAU(1MG)(PSU)(5MU)GAAA AAUUAGCGGA UGACUUGUGG CUGGGGGUGA AAGGCCA AU CAAACCGGGA GAUAGCUGGU UCUCCCCGAA AGCUAUUUAG GUAGCGCCUC GUGAAUUCAU CUCCGGGGGU AGAGCACU G UUUCGGCAAG GGGGUCAACC CGACUUACCA ACCCGAUGCA AACUGCGAAU ACCGGAGAAU GUUAUCACGG GAGACACAC GGCGGG(PSU)GCU AACGUCCGUC GUGAAGAGGG AAACAACCCA GACCGCCAGC UAAGGUCCCA AAGUCAUGGU UAAGUG GGA AACGAUGUGG GAAGGCCCAG ACAGCCAGGA UGUUGGCUUA GAAGCAGCCA UCAUUUAAAG AAAGCGUAAU AGCUCAC UG GUCGAGUCGG CCUGCGCGGA AGAUGUAACG GGGCUAAACC AUGCACCGAA GCUGCGGCAG CGACGCUUAU GCGUUGUU G GGUAGGGGAG CGUUCUGUAA GCCUGCGAAG GUGUGCUGUG AGGCAUGCUG GAGGUAUCAG AAGUGCGAAU GCUGACAUA AGUAACGAUA AAGCGGGUGA AAAGCCCGCU CGCCGGAAGA CCAAGGGUUC CUGUCCAACG UUAAUCGGGG CAGGGUGAGU CGACCCCUA AGGCGAGGCC GAAAGGCGUA GUCGAUGGGA AACAGGUUAA UAUUCCUGUA CUUGGUGUUA CUGCGAAGGG G GGACGGAG AAGGCUAUGU UGGCCGGGCG ACGGUUGUCC CGGUUUAAGC GUGUAGGCUG GUUUUCCAGG CAAAUCCGGA AA AUCAAGG CUGAGGCGUG AUGACGAGGC ACUACGGUGC UGAAGCAACA AAUGCCCUGC UUCCAGGAAA AGCCUCUAAG CAU CAGGUA ACAUCAAAUC GUACCCCAAA CCGACAC(6MZ)GG UGGUCAGGUA GAGAAUACCA AGGCGCUUGA GAGAACUCGG GUGAAGGAA CUAGGCAAAA UGGUGCCGUA ACUUCGGGAG AAGGCACGCU GAUAUGUAGG UGAGGUCCCU CGCGGAUGGA G CUGAAAUC AGUCGAAGAU ACCAGCUGGC UGCAACUGUU UAUUAAAAAC ACAGCACUGU GCAAACACGA AAGUGGACGU AU ACGGUGU GACGCCU(2MG)CC CGGUGCCGGA AGGUUAAUUG AUGGGGUUAG CGCAAGCGAA GCUCUUGAUC GAAGCCCCG GUAAACGGCG GCCG(PSU)AAC(3TD)A (PSU)AACGGUCCU AAGGUAGCGA AA(5MU)UCCUUGU CGGGUAAGUU CCGA C(5MC)UGC ACGAAUGGCG UAAUGAUGGC CAGGCUGUCU CCACCCGAGA CUCAGUGAAA UUGAACUCGC UGUG(6MZ)AG AU GCAGUGUACC CGCGGCAAGA CGGAAAGACC CCGU(G7M)AACCU UUACUAUAGC UUGACACUGA ACAUUGAGCC UUGA UGUGU AGGAUAGGUG GGAGGCUUUG AAGUGUGGAC GCCAGUCUGC AUGGAGCCGA CCUUGAAAUA CCACCCUUUA AUGUU UGAU GUUCUAACGU UGACCCGUAA UCCGGGUUGC GGACAGUGUC UGGUGGGUAG UUUGACUG(OMG)G GCGGUCUCCU CC UAAAGAG UAACGGAGGA GCACGAAGGU UGGCUAAUCC UGGUCGGACA UCAGGAGGUU AGUGCAAUGG CAUAAGCCAG CUU GACUGC GAGCGUGACG GCGCGAGCAG GUGCGAAAGC AGGUCAUAGU GAUCCGGUGG UUCUGAAUGG AAGGGCCAUC GCUC AACGG AUAAAAGGUA CUCCG(2MG)GGAU AACAGGC(PSU)GA UACCGCCCAA GAGUUCAUAU CGACGGCGGU GUUUGGC A(OMC) CUCG(2MA)(PSU)GUCG GCUCAUCACA UCCUGGGGCU GAAGUAGGUC CCAAGGGUAU GGC(OMU)GUUCGC CA UUUAAAG UGGUACGCGA GC(PSU)GGGUUUA GAACGUCGUG AGACAGU(PSU)CG GUCCCUAUCU GCCGUGGGCG CUGGA GAAC UGAGGGGGGC UGCUCCUAGU ACGAGAGGAC CGGAGUGGAC GCAUCACUGG UGUUCGGGUU GUCAUGCCAA UGGCAC UGC CCGGUAGCUA AAUGCGGAAG AGAUAAGUGC UGAAAGCAUC UAAGCACGAA ACUUGCCCCG AGAUGAGUUC UCCCUGA CC CUUUAAGGGU CCUGAAGGAA CGUUGAAGAC GACGACGUUG AUAGGCCGGG UGUGUAAGCG CAGCGAUGCG UUGAGCUA A CCGGUACUAA UGAACCGUGA GGCUUAACCU U

Macromolecule #27: 16S ribosomal RNA

• Macromolecule: Name: 16S ribosomal RNA / type: rna / ID: 27 / Number of copies: 1
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 497.405969 KDa

Sequence

String:

AAAUUGAAGA GUUUGAUCAU GGCUCAGAUU GAACGCUGGC GGCAGGCCUA ACACAUGCAA GUCGAACGGU AACAGGAAGA AGCUUGCUU CUUUGCUGAC GAGUGGCGGA CGGGUGAGUA AUGUCUGGGA AACUGCCUGA UGGAGGGGGA UAACUACUGG A AACGGUAG CUAAUACCGC AUAACGUCGC AAGACCAAAG AGGGGGACCU UCGGGCCUCU UGCCAUCGGA UGUGCCCAGA UG GGAUUAG CUAGUAGGUG GGGUAACGGC UCACCUAGGC GACGAUCCCU AGCUGGUCUG AGAGGAUGAC CAGCCACACU GGA ACUGAG ACACGGUCCA GACUCCUACG GGAGGCAGCA GUGGGGAAUA UUGCACAAUG GGCGCAAGCC UGAUGCAGCC AUGC CGCGU GUAUGAAGAA GGCCUUCGGG UUGUAAAGUA CUUUCAGCGG GGAGGAAGGG AGUAAAGUUA AUACCUUUGC UCAUU GACG UUACCCGCAG AAGAAGCACC GGCUAACUCC G(PSU)GCCAGCAG CC(7MG)CGGUAAU ACGGAGGGUG CAAGCGUU A AUCGGAAUUA CUGGGCGUAA AGCGCACGCA GGCGGUUUGU UAAGUCAGAU GUGAAAUCCC CGGGCUCAAC CUGGGAACU GCAUCUGAUA CUGGCAAGCU UGAGUCUCGU AGAGGGGGGU AGAAUUCCAG GUGUAGCGGU GAAAUGCGUA GAGAUCUGGA GGAAUACCG GUGGCGAAGG CGGCCCCCUG GACGAAGACU GACGCUCAGG UGCGAAAGCG UGGGGAGCAA ACAGGAUUAG A UACCCUGG UAGUCCACGC CGUAAACGAU GUCGACUUGG AGGUUGUGCC CUUGAGGCGU GGCUUCCGGA GCUAACGCGU UA AGUCGAC CGCCUGGGGA GUACGGCCGC AAGGUUAAAA CUCAAAUGAA UUGACGGGGG CCCGCACAAG CGGUGGAGCA UGU GGUUUA AUUCGAU(2MG)(5MC)A ACGCGAAGAA CCUUACCUGG UCUUGACAUC CACGGAAGUU UUCAGAGAUG AGAAUG UGC CUUCGGGAAC CGUGAGACAG GUGCUGCAUG GCUGUCGUCA GCUCGUGUUG UGAAAUGUUG GGUUAAGUCC CGCAACG AG CGCAACCCUU AUCCUUUGUU GCCAGCGGUC CGGCCGGGAA CUCAAAGGAG ACUGCCAGUG AUAAACUGGA GGAAGGUG G GGAUGACGUC AAGUCAUCAU G(2MG)CCCUUACG ACCAGGGCUA CACACGUGCU ACAAUGGCGC AUACAAAGAG AAGCG ACCU CGCGAGAGCA AGCGGACCUC AUAAAGUGCG UCGUAGUCCG GAUUGGAGUC UGCAACUCGA CUCCAUGAAG UCGGAA UCG CUAGUAAUCG UGGAUCAGAA UGCCACGGUG AAUACGUUCC CGGGCCUUGU ACACACCG(4OC)C CGU(5MC)ACACC AUGGGAGUGG GUUGCAAAAG AAGUAGGUAG CUUAACCUUC GGGAGGGCGC UUACCACUUU GUGAUUCAUG ACUGGGGUGA AGUCG(UR3)AAC AAGGUAACCG UAGG(2MG)G(MA6)(MA6)CC UGCGGUUGGA UCA

GENBANK: GENBANK: CP032808.1

Macromolecule #28: 5S ribosomal RNA

• Macromolecule: Name: 5S ribosomal RNA / type: rna / ID: 28 / Number of copies: 1
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 38.79009 KDa

Sequence

String:

UGCCUGGCGG CCGUAGCGCG GUGGUCCCAC CUGACCCCAU GCCGAACUCA GAAGUGAAAC GCCGUAGCGC CGAUGGUAGU GUGGGGUCU CCCCAUGCGA GAGUAGGGAA CUGCCAGGCA U

GENBANK: GENBANK: CP035706.1

Macromolecule #29: fMet-NH-tRNA(fMet)

• Macromolecule: Name: fMet-NH-tRNA(fMet) / type: rna / ID: 29 / Number of copies: 1
• Source (natural): Organism: Escherichia coli (E. coli)
• Molecular weight: Theoretical: 24.806922 KDa

Sequence

String:

CGCGGGGU(4SU)G AGCAGCCUGG (H2U)AGCUCGUCG GG(4OC)UCAUAAC CCGAAGAUCG UCGG(5MU)(PSU)CAAA UCCGGCCCC CGCAACC(8AN)

GENBANK: GENBANK: CP025520.1

Macromolecule #56: mRNA

• Macromolecule: Name: mRNA / type: rna / ID: 56 ; Details: EMCV frameshift site flanked by the bacterial 5' UTR with Shine-Dalgarno sequence and 18 nt downstream region of the putative structure; Number of copies: 1
• Source (natural): Organism: Encephalomyocarditis virus
• Molecular weight: Theoretical: 37.684395 KDa

Sequence

String:

GGGAAUUCAA AAAUUGUUAA GAAUUAAGGA GAUAUACAUA UGGAGGUUUU UAUCACUCAA GGAGCGGCAG UGUCAUCAAU GGCUCAAAC CCUACUGCCG AACGACUUGG CCAGAUCU

Macromolecule #58: MAGNESIUM ION

• Macromolecule: Name: MAGNESIUM ION / type: ligand / ID: 58 / Number of copies: 437 / Formula: MG
• Molecular weight: Theoretical: 24.305 Da

Macromolecule #59: N-FORMYLMETHIONINE

• Macromolecule: Name: N-FORMYLMETHIONINE / type: ligand / ID: 59 / Number of copies: 1 / Formula: FME
• Molecular weight: Theoretical: 177.221 Da

Chemical component information

ChemComp-FME:
N-FORMYLMETHIONINE

Macromolecule #60: ZINC ION

• Macromolecule: Name: ZINC ION / type: ligand / ID: 60 / Number of copies: 2 / Formula: ZN
• Molecular weight: Theoretical: 65.409 Da

Macromolecule #61: water

• Macromolecule: Name: water / type: ligand / ID: 61 / Number of copies: 2 / Formula: HOH
• Molecular weight: Theoretical: 18.015 Da

Chemical component information

ChemComp-HOH:
WATER

Experimental details

Structure determination

• Method: cryo EM
• Processing: single particle reconstruction
• Aggregation state: particle

Sample preparation

Buffer

pH: 7.5
Component:

Concentration	Name
20.0 mM	HEPES
100.0 mM	potassium acetate
2.2 mM	magnesium chloride
2.0 mM	dithiothreitol
5.0 mM	Tris-HCl
7.0 mM	ammonium acetate
3.0 mM	potassium chloride

Details: Initiated 70S ribosomes in 50 mM Tris-HCl pH 7.5, 70 mM NH4Cl, 30 mM KCl, 7 mM MgCl2 were diluted tenfold into 20 mM HEPES pH 7.5, 100 mM potassium acetate, 1.5 mM MgCl2, 2.0 mM DTT. 2A protein was dialysed (3K MWCO, 277K, 16 h) into the same buffer. Crosslinking reactions of 50 microliters comprising 75 nM ribosomes, 3.0 micromolar 2A and 2.0 mM bis(sulfosuccinimidyl)suberate (BS3) were performed on ice (30 min) immediately prior to grid preparation.

• Grid: Model: Quantifoil R2/2 / Material: COPPER / Mesh: 400 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 30 sec. / Pretreatment - Atmosphere: AIR
• Vitrification: Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV; Details: Quantifoil R 2/2 400-mesh copper supports were coated with an additional ~ 60 angstrom layer of amorphous, evaporated carbon by flotation and thoroughly dried before use. Grids were made hydrophilic by glow-discharge in air for 30 s. Three microliters of crosslinking reaction was applied to grids which were then blotted for 4.5 s and vitrified by plunging into liquid ethane using a Vitrobot MK IV (FEI) at 277K, 100% relative humidity..
• Details: 75 nM ribosomes, 3.0 micromolar 2A crosslinked with 2.0 mM bis(sulfosuccinimidyl)suberate (BS3)

Electron microscopy

• Microscope: FEI TITAN KRIOS
• Image recording: Film or detector model: FEI FALCON III (4k x 4k) / Detector mode: INTEGRATING / Number grids imaged: 1 / Number real images: 5730 / Average exposure time: 0.59 sec. / Average electron dose: 54.4 e/Ų; Details: Images were collected as 23-frame movies. Autofocus every 10 micrometres, no drift measurement, 7 sec delay after stage shift and 3 sec delay after image shift
• Electron beam: Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
• Electron optics: C2 aperture diameter: 100.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 3.0 µm / Nominal defocus min: 1.2 µm / Nominal magnification: 75000
• Sample stage: Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
• Experimental equipment: Model: Titan Krios / Image courtesy: FEI Company

Image processing

• Details: Movie frames were aligned and a dose-weighted average calculated with MotionCor2
• Particle selection: Number selected: 820475 ; Details: Reference-free autopicking of particles was performed using the Laplacian-of-Gaussian function in Relion (200 - 250 angstrom diameter)

Startup model

Type of model: EMDB MAP
EMDB ID:

3493

Details: Map was low-pass filtered to 80 angstroms resolution

• Final reconstruction: Number classes used: 1 / Applied symmetry - Point group: C₁ (asymmetric) / Algorithm: FOURIER SPACE / Resolution.type: BY AUTHOR / Resolution: 2.66 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 3.1) / Number images used: 120749
• Initial angle assignment: Type: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 3.1)
• Final angle assignment: Type: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 3.1)
• Final 3D classification: Number classes: 6 / Avg.num./class: 20124 / Software - Name: RELION (ver. 3.1); Details: Focussed classification with signal subtraction and local angular searches was performed to separate particles based on 2A occupancy at the factor binding site

Atomic model buiding 1

Initial model

PDB ID:

5mdz

Chain - Source name: PDB / Chain - Initial model type: experimental model

• Refinement: Space: REAL / Protocol: FLEXIBLE FIT / Target criteria: Correlation coefficient

Output model

PDB-7nwt:
Initiated 70S ribosome in complex with 2A protein from encephalomyocarditis virus (EMCV)