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- EMDB-9258: The structure of the Plasmodium falciparum 20S proteasome. -

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Basic information

Entry
Database: EMDB / ID: EMD-9258
TitleThe structure of the Plasmodium falciparum 20S proteasome.
Map dataSharpened map used for refinement
Sample
  • Complex: 20S proteasome/PA28 complex.
    • Protein or peptide: x 14 types
Keywordsproteasome / protease / HYDROLASE
Function / homology
Function and homology information


Cross-presentation of soluble exogenous antigens (endosomes) / Orc1 removal from chromatin / CDK-mediated phosphorylation and removal of Cdc6 / FBXL7 down-regulates AURKA during mitotic entry and in early mitosis / KEAP1-NFE2L2 pathway / UCH proteinases / Ub-specific processing proteases / Neddylation / Antigen processing: Ubiquitination & Proteasome degradation / MAPK6/MAPK4 signaling ...Cross-presentation of soluble exogenous antigens (endosomes) / Orc1 removal from chromatin / CDK-mediated phosphorylation and removal of Cdc6 / FBXL7 down-regulates AURKA during mitotic entry and in early mitosis / KEAP1-NFE2L2 pathway / UCH proteinases / Ub-specific processing proteases / Neddylation / Antigen processing: Ubiquitination & Proteasome degradation / MAPK6/MAPK4 signaling / ABC-family proteins mediated transport / AUF1 (hnRNP D0) binds and destabilizes mRNA / Neutrophil degranulation / proteasome core complex / proteasomal ubiquitin-independent protein catabolic process / proteasome endopeptidase complex / proteasome core complex, beta-subunit complex / proteasome core complex, alpha-subunit complex / threonine-type endopeptidase activity / proteasomal protein catabolic process / ubiquitin-dependent protein catabolic process / proteasome-mediated ubiquitin-dependent protein catabolic process / endopeptidase activity / hydrolase activity / nucleus / cytosol / cytoplasm
Similarity search - Function
Proteasome subunit beta 1 / Proteasome subunit alpha 1 / Proteasome subunit beta 4 / Proteasome subunit beta 2 / Proteasome beta 3 subunit / Proteasome subunit alpha6 / Proteasome subunit alpha5 / Proteasome beta-type subunits signature. / Peptidase T1A, proteasome beta-subunit / Proteasome beta-type subunit, conserved site ...Proteasome subunit beta 1 / Proteasome subunit alpha 1 / Proteasome subunit beta 4 / Proteasome subunit beta 2 / Proteasome beta 3 subunit / Proteasome subunit alpha6 / Proteasome subunit alpha5 / Proteasome beta-type subunits signature. / Peptidase T1A, proteasome beta-subunit / Proteasome beta-type subunit, conserved site / Proteasome subunit A N-terminal signature / Proteasome alpha-type subunits signature. / Proteasome alpha-subunit, N-terminal domain / Proteasome subunit A N-terminal signature Add an annotation / : / Proteasome alpha-type subunit / Proteasome alpha-type subunit profile. / Proteasome B-type subunit / Proteasome beta-type subunit profile. / Proteasome subunit / Proteasome, subunit alpha/beta / Nucleophile aminohydrolases, N-terminal
Similarity search - Domain/homology
Proteasome subunit beta / Proteasome subunit alpha type-2, putative / Proteasome subunit alpha type-3, putative / Proteasome subunit beta / Proteasome subunit beta type-6, putative / Proteasome subunit beta / Proteasome subunit beta / Proteasome subunit alpha type-6, putative / Proteasome subunit alpha type / Proteasome subunit alpha type ...Proteasome subunit beta / Proteasome subunit alpha type-2, putative / Proteasome subunit alpha type-3, putative / Proteasome subunit beta / Proteasome subunit beta type-6, putative / Proteasome subunit beta / Proteasome subunit beta / Proteasome subunit alpha type-6, putative / Proteasome subunit alpha type / Proteasome subunit alpha type / Proteasome subunit alpha type / Proteasome subunit beta / Proteasome subunit alpha type-1, putative / Proteasome subunit beta
Similarity search - Component
Biological speciesPlasmodium falciparum 3D7 (eukaryote) / Plasmodium falciparum (isolate 3D7) (eukaryote)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.6 Å
AuthorsHanssen E / Xie SC / Leis A / Metcalfe RD / Gillett DL / Tilley L / Griffin MDW
Funding support Australia, 2 items
OrganizationGrant numberCountry
National Health and Medical Research Council (NHMRC, Australia) Australia
Australian Research Council (ARC) Australia
CitationJournal: Nat Microbiol / Year: 2019
Title: The structure of the PA28-20S proteasome complex from Plasmodium falciparum and implications for proteostasis.
Authors: Stanley C Xie / Riley D Metcalfe / Eric Hanssen / Tuo Yang / David L Gillett / Andrew P Leis / Craig J Morton / Michael J Kuiper / Michael W Parker / Natalie J Spillman / Wilson Wong / ...Authors: Stanley C Xie / Riley D Metcalfe / Eric Hanssen / Tuo Yang / David L Gillett / Andrew P Leis / Craig J Morton / Michael J Kuiper / Michael W Parker / Natalie J Spillman / Wilson Wong / Christopher Tsu / Lawrence R Dick / Michael D W Griffin / Leann Tilley /
Abstract: The activity of the proteasome 20S catalytic core is regulated by protein complexes that bind to one or both ends. The PA28 regulator stimulates 20S proteasome peptidase activity in vitro, but its ...The activity of the proteasome 20S catalytic core is regulated by protein complexes that bind to one or both ends. The PA28 regulator stimulates 20S proteasome peptidase activity in vitro, but its role in vivo remains unclear. Here, we show that genetic deletion of the PA28 regulator from Plasmodium falciparum (Pf) renders malaria parasites more sensitive to the antimalarial drug dihydroartemisinin, indicating that PA28 may play a role in protection against proteotoxic stress. The crystal structure of PfPA28 reveals a bell-shaped molecule with an inner pore that has a strong segregation of charges. Small-angle X-ray scattering shows that disordered loops, which are not resolved in the crystal structure, extend from the PfPA28 heptamer and surround the pore. Using single particle cryo-electron microscopy, we solved the structure of Pf20S in complex with one and two regulatory PfPA28 caps at resolutions of 3.9 and 3.8 Å, respectively. PfPA28 binds Pf20S asymmetrically, strongly engaging subunits on only one side of the core. PfPA28 undergoes rigid body motions relative to Pf20S. Molecular dynamics simulations support conformational flexibility and a leaky interface. We propose lateral transfer of short peptides through the dynamic interface as a mechanism facilitating the release of proteasome degradation products.
History
DepositionOct 23, 2018-
Header (metadata) releaseNov 7, 2018-
Map releaseAug 7, 2019-
UpdateMar 13, 2024-
Current statusMar 13, 2024Processing site: RCSB / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 5
  • Imaged by UCSF Chimera
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  • Surface view colored by cylindrical radius
  • Surface level: 5
  • Imaged by UCSF Chimera
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  • Surface view with fitted model
  • Atomic models: PDB-6muw
  • Surface level: 5
  • Imaged by UCSF Chimera
  • Download
Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

emd_9258.png

Downloads & links

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Map

FileDownload / File: emd_9258.map.gz / Format: CCP4 / Size: 155.3 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationSharpened map used for refinement
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesX (Sec.)Y (Row.)Z (Col.)
1.31 Å/pix.
x 344 pix.
= 450.64 Å		1.31 Å/pix.
x 344 pix.
= 450.64 Å		1.31 Å/pix.
x 344 pix.
= 450.64 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 1.31 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 5.0 / Movie #1: 5
Minimum - Maximum-18.409132 - 32.030929999999998
Average (Standard dev.)-0.000000000002087 (±1.0)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderZYX
Origin000
Dimensions344344344
Spacing344344344
CellA=B=C: 450.63998 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z1.311.311.31
M x/y/z344344344
origin x/y/z0.0000.0000.000
length x/y/z450.640450.640450.640
α/β/γ90.00090.00090.000
start NX/NY/NZ000
NX/NY/NZ344344344
MAP C/R/S321
start NC/NR/NS000
NC/NR/NS344344344
D min/max/mean-18.40932.031-0.000

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Supplemental data

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Additional map: Initial unsharpened map

Fileemd_9258_additional.map
AnnotationInitial unsharpened map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: Half-map 2

Fileemd_9258_half_map_1.map
AnnotationHalf-map 2
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: Half-map 1

Fileemd_9258_half_map_2.map
AnnotationHalf-map 1
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Sample components

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Entire : 20S proteasome/PA28 complex.

EntireName: 20S proteasome/PA28 complex.
Components
  • Complex: 20S proteasome/PA28 complex.
    • Protein or peptide: 20S proteasome alpha-1 subunit
    • Protein or peptide: 20S proteasome alpha-2 subunit
    • Protein or peptide: 20S proteasome alpha-3 subunit
    • Protein or peptide: 20S proteasome alpha-4 subunit
    • Protein or peptide: 20S proteasome alpha-5 subunit
    • Protein or peptide: 20S proteasome alpha-6 subunit
    • Protein or peptide: 20S proteasome alpha-7 subunit
    • Protein or peptide: 20S proteasome beta-1 subunit
    • Protein or peptide: 20S proteasome beta-2 subunit
    • Protein or peptide: 20S proteasome beta-3 subunit
    • Protein or peptide: 20S proteasome beta-4 subunit
    • Protein or peptide: 20S proteasome beta-5 subunit
    • Protein or peptide: 20S proteasome beta-6 subunit
    • Protein or peptide: 20S proteasome beta-7 subunit

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Supramolecule #1: 20S proteasome/PA28 complex.

SupramoleculeName: 20S proteasome/PA28 complex. / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all
Details: Sample was a mixture of unbound 20S proteasome, 20S proteasome in complex with one PA28 cap, and 20S proteasome in complex with two PA28 caps.
Source (natural)Organism: Plasmodium falciparum 3D7 (eukaryote)
Molecular weightTheoretical: 760 KDa

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Macromolecule #1: 20S proteasome alpha-1 subunit

MacromoleculeName: 20S proteasome alpha-1 subunit / type: protein_or_peptide / ID: 1 / Number of copies: 2 / Enantiomer: LEVO / EC number: proteasome endopeptidase complex
Source (natural)Organism: Plasmodium falciparum (isolate 3D7) (eukaryote) / Strain: isolate 3D7
Molecular weightTheoretical: 29.531656 KDa
SequenceString: MVRPSQSMYD RHLTIFSPDG NLYQIEYAIK AVKNTNITSV GVKGENCAVI ISQKKMATQY ISQDKLLDYN NITNIYNITD EIGCSMVGM PGDCLSMVYK ARSEASEFLY SNGYNVNAET LCRNICDKIQ VYTQHAYMRL HACSGMIIGI DENNKPELFK F DPSGFCAG ...String:
MVRPSQSMYD RHLTIFSPDG NLYQIEYAIK AVKNTNITSV GVKGENCAVI ISQKKMATQY ISQDKLLDYN NITNIYNITD EIGCSMVGM PGDCLSMVYK ARSEASEFLY SNGYNVNAET LCRNICDKIQ VYTQHAYMRL HACSGMIIGI DENNKPELFK F DPSGFCAG YRACVIGNKE QESISVLERL LEKRKKKIQQ ETIDEDIRNT TILAIEALQT ILAFDLKASE IEVAIVSTKN RN FTQISEK EIDNYLTYIA ERD

UniProtKB: Proteasome subunit alpha type-6, putative

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Macromolecule #2: 20S proteasome alpha-2 subunit

MacromoleculeName: 20S proteasome alpha-2 subunit / type: protein_or_peptide / ID: 2 / Number of copies: 2 / Enantiomer: LEVO / EC number: proteasome endopeptidase complex
Source (natural)Organism: Plasmodium falciparum (isolate 3D7) (eukaryote) / Strain: isolate 3D7
Molecular weightTheoretical: 26.556391 KDa
SequenceString: MADGEYSFSL TTFSPTGKLV QIEYALNRVS SSSPALGIRA KNGVIIATEK KSPNELIEEN SIFKIQQISE HIGIVYAGMP GDFRVLLKR ARKEAIRYSL QYGSEILVKE LVKIIASIVQ EFTQTGGVRP FGLSLLICGV DVYGYHLYQI DPSGCYFNWM A TCVGKDYQ ...String:
MADGEYSFSL TTFSPTGKLV QIEYALNRVS SSSPALGIRA KNGVIIATEK KSPNELIEEN SIFKIQQISE HIGIVYAGMP GDFRVLLKR ARKEAIRYSL QYGSEILVKE LVKIIASIVQ EFTQTGGVRP FGLSLLICGV DVYGYHLYQI DPSGCYFNWM A TCVGKDYQ NNMSFLEKRY NKDIEIEDAI HTAILTLKES YEGVLNEKNI EIGVAYDNKP FKILTQNEIK DYLIEIE

UniProtKB: Proteasome subunit alpha type-2, putative

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Macromolecule #3: 20S proteasome alpha-3 subunit

MacromoleculeName: 20S proteasome alpha-3 subunit / type: protein_or_peptide / ID: 3 / Number of copies: 2 / Enantiomer: LEVO / EC number: proteasome endopeptidase complex
Source (natural)Organism: Plasmodium falciparum (isolate 3D7) (eukaryote) / Strain: isolate 3D7
Molecular weightTheoretical: 27.977664 KDa
SequenceString: MARRYDSRTT TFSPEGRLYQ VEYALEAINN ASITIGLITK DGVILGADKV FISKLIDKAN NYEKIYKIDK HIFCGVAGLN ADANILINQ SRLYAQRYLY NYNEVQPVSQ LVVQICDIKQ SYTQYGGLRP YGVSFLIGGY DTKDGYQLYH TDPSGNYSGW F ATAIGTNN ...String:
MARRYDSRTT TFSPEGRLYQ VEYALEAINN ASITIGLITK DGVILGADKV FISKLIDKAN NYEKIYKIDK HIFCGVAGLN ADANILINQ SRLYAQRYLY NYNEVQPVSQ LVVQICDIKQ SYTQYGGLRP YGVSFLIGGY DTKDGYQLYH TDPSGNYSGW F ATAIGTNN LTASSVLKQE WKNDMTLEEG LLLALKTLAK STDTEIPKSE KIELAYLTNK DGEVYQKYLT EKEIEELIKL YT QKYIKE

UniProtKB: Proteasome subunit alpha type

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Macromolecule #4: 20S proteasome alpha-4 subunit

MacromoleculeName: 20S proteasome alpha-4 subunit / type: protein_or_peptide / ID: 4 / Number of copies: 2 / Enantiomer: LEVO / EC number: proteasome endopeptidase complex
Source (natural)Organism: Plasmodium falciparum (isolate 3D7) (eukaryote) / Strain: isolate 3D7
Molecular weightTheoretical: 27.263285 KDa
SequenceString: MSYDRAITVF SPDGHLLQVE HALEAVKKGG CAVAIKSSNF AVLAVEKKNI PKLQNPKTTE KLIKLDEHNC LAFAGLNADA RVLVNKTRL ECQRYYLNMD EPAPVDYIAK YVAKVQQKFT HRGGVRPFGI ATLIAGFKNN KEICIYQTEP SGIYAAWKAQ A IGKNAKIV ...String:
MSYDRAITVF SPDGHLLQVE HALEAVKKGG CAVAIKSSNF AVLAVEKKNI PKLQNPKTTE KLIKLDEHNC LAFAGLNADA RVLVNKTRL ECQRYYLNMD EPAPVDYIAK YVAKVQQKFT HRGGVRPFGI ATLIAGFKNN KEICIYQTEP SGIYAAWKAQ A IGKNAKIV QEFLEKNYQE NMEQKDCIFL ALKAIFEVVE LSSKNVEVAL LTEKDLTFIE EQEINSMVEL IDQERTKNNE QN E

UniProtKB: Proteasome subunit alpha type

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Macromolecule #5: 20S proteasome alpha-5 subunit

MacromoleculeName: 20S proteasome alpha-5 subunit / type: protein_or_peptide / ID: 5 / Number of copies: 2 / Enantiomer: LEVO / EC number: proteasome endopeptidase complex
Source (natural)Organism: Plasmodium falciparum (isolate 3D7) (eukaryote) / Strain: isolate 3D7
Molecular weightTheoretical: 28.417367 KDa
SequenceString: MFSTRSEYDR GVNTFSPEGR LFQVEYALGA IKLGSTAVGI CVNDGVILAS ERRISSTLIE KDSVEKLLSI DDHIGCAMSG LMADARTLI DYARVECNHY KFIYNENINI KSCVELISEL ALDFSNLSDS KRKKIMSRPF GVALLIGGVD KNGPCLWYTE P SGTNTRFS ...String:
MFSTRSEYDR GVNTFSPEGR LFQVEYALGA IKLGSTAVGI CVNDGVILAS ERRISSTLIE KDSVEKLLSI DDHIGCAMSG LMADARTLI DYARVECNHY KFIYNENINI KSCVELISEL ALDFSNLSDS KRKKIMSRPF GVALLIGGVD KNGPCLWYTE P SGTNTRFS AASIGSAQEG AELLLQENYK KDMTFEQAEI LALTVLRQVM EDKLSTSNVE ICAIKKSDQT FYKYNTDDIS RI IDVLPSP VYPTIDMTA

UniProtKB: Proteasome subunit alpha type

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Macromolecule #6: 20S proteasome alpha-6 subunit

MacromoleculeName: 20S proteasome alpha-6 subunit / type: protein_or_peptide / ID: 6 / Number of copies: 2 / Enantiomer: LEVO / EC number: proteasome endopeptidase complex
Source (natural)Organism: Plasmodium falciparum (isolate 3D7) (eukaryote) / Strain: isolate 3D7
Molecular weightTheoretical: 28.871697 KDa
SequenceString: MYRNLYDTDN IIYSPEGRLY QVEYASEAIK QGTCAVAIKS KDYVVVSGLK KCISKLSFPQ EKIFKIDDYI GISMSGITSD AKVLTKFMQ NECLSHKFLY NENINIESLV RSVADKYQKN TQKSSKRAFG VGLMIAAYHN EPCIFETRPN GSYFEYDALS F GARSHASK ...String:
MYRNLYDTDN IIYSPEGRLY QVEYASEAIK QGTCAVAIKS KDYVVVSGLK KCISKLSFPQ EKIFKIDDYI GISMSGITSD AKVLTKFMQ NECLSHKFLY NENINIESLV RSVADKYQKN TQKSSKRAFG VGLMIAAYHN EPCIFETRPN GSYFEYDALS F GARSHASK TYLEKNLHLF EECSLEELIL HCLKALKCSL SSESELTISN TALAVVGKNH PWQEISSLQL EEYLSKVKMD AE QEQVEEN VQNEANE

UniProtKB: Proteasome subunit alpha type-1, putative

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Macromolecule #7: 20S proteasome alpha-7 subunit

MacromoleculeName: 20S proteasome alpha-7 subunit / type: protein_or_peptide / ID: 7 / Number of copies: 2 / Enantiomer: LEVO / EC number: proteasome endopeptidase complex
Source (natural)Organism: Plasmodium falciparum (isolate 3D7) (eukaryote) / Strain: isolate 3D7
Molecular weightTheoretical: 29.324295 KDa
SequenceString: MAGLSAGYDL SVSTFSPDGR LYQVEYIYKS INNNNTALCL ECKDGIICCC INSNMDKNKM IKKNSYNRIY HVNNNIIITY SGFDGDARN IIDRARSEAN TYYYNFHTNI PLHILVNRIS LYIHAYTLYW HMRPFAASII ISSFNEKDKG DIYCIEPNGA C YKYSGIVI ...String:
MAGLSAGYDL SVSTFSPDGR LYQVEYIYKS INNNNTALCL ECKDGIICCC INSNMDKNKM IKKNSYNRIY HVNNNIIITY SGFDGDARN IIDRARSEAN TYYYNFHTNI PLHILVNRIS LYIHAYTLYW HMRPFAASII ISSFNEKDKG DIYCIEPNGA C YKYSGIVI GKNKEMFKTE IEKKDYKDIN VRDAIEDIYK FILTSDDHMN KNNLQNLVNF SWICKESSYE FQNIHEEILT PA LNKAVEY IEKLN

UniProtKB: Proteasome subunit alpha type-3, putative

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Macromolecule #8: 20S proteasome beta-1 subunit

MacromoleculeName: 20S proteasome beta-1 subunit / type: protein_or_peptide / ID: 8 / Number of copies: 2 / Enantiomer: LEVO / EC number: proteasome endopeptidase complex
Source (natural)Organism: Plasmodium falciparum (isolate 3D7) (eukaryote) / Strain: isolate 3D7
Molecular weightTheoretical: 29.143936 KDa
SequenceString: TTIIGIIYDN GVMLACDSRT SSGTFISNKC SRKINRINEN LYVCRSGASA HSQKIIEIIK HYCVSMKNEN RKKGRFHEGE TIYDETTYD EEIDIDSINY LDYNNNNDNN LVTKNKYFYE DKFNDYNPLV ENVAHITKKI IYTNNNFLSC ALIFGGYDKI K KQQLYAVN ...String:
TTIIGIIYDN GVMLACDSRT SSGTFISNKC SRKINRINEN LYVCRSGASA HSQKIIEIIK HYCVSMKNEN RKKGRFHEGE TIYDETTYD EEIDIDSINY LDYNNNNDNN LVTKNKYFYE DKFNDYNPLV ENVAHITKKI IYTNNNFLSC ALIFGGYDKI K KQQLYAVN LNGSIIEKHD FAVSGSGSIY IQSYLQDKYK KFMTKKECFN LILNCVKYAM HNDNSSGGLI RIVNITKSFV EE FTVVNTQ MNFQY

UniProtKB: Proteasome subunit beta type-6, putative

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Macromolecule #9: 20S proteasome beta-2 subunit

MacromoleculeName: 20S proteasome beta-2 subunit / type: protein_or_peptide / ID: 9 / Number of copies: 2 / Enantiomer: LEVO / EC number: proteasome endopeptidase complex
Source (natural)Organism: Plasmodium falciparum (isolate 3D7) (eukaryote) / Strain: isolate 3D7
Molecular weightTheoretical: 25.104885 KDa
SequenceString: TTICGLVCQN AVILGADTRA TEGPIVADKN CSKLHYISKN IWCAGAGVAG DLEHTTLWLQ HNVELHRLNT NTQPRVSMCV SRLTQELFK YQGYKVCAIV LGGVDVNGPQ LYGIHPHGSS CLLPFTALGS GSLNAMAVLE AKYRDNMTIE EGKNLVCEAI C AGIFNDLG ...String:
TTICGLVCQN AVILGADTRA TEGPIVADKN CSKLHYISKN IWCAGAGVAG DLEHTTLWLQ HNVELHRLNT NTQPRVSMCV SRLTQELFK YQGYKVCAIV LGGVDVNGPQ LYGIHPHGSS CLLPFTALGS GSLNAMAVLE AKYRDNMTIE EGKNLVCEAI C AGIFNDLG SGGNVDICVI TKDSYQHIRP YKEPNMRLYH LPHPTIYPKG TTPILSEKIE YIKKFISVED A

UniProtKB: Proteasome subunit beta

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Macromolecule #10: 20S proteasome beta-3 subunit

MacromoleculeName: 20S proteasome beta-3 subunit / type: protein_or_peptide / ID: 10 / Number of copies: 2 / Enantiomer: LEVO / EC number: proteasome endopeptidase complex
Source (natural)Organism: Plasmodium falciparum (isolate 3D7) (eukaryote) / Strain: isolate 3D7
Molecular weightTheoretical: 24.533131 KDa
SequenceString: MGSIYNYNGG CVLGMSGSNC VAIACDLRLG ANTFTTVSTK FSKIFKMNNN VYVGLSGLAT DIQTLYEILR YRVNLYEVRQ DAEMDVECF ANMLSSILYS NRFSPYFVNP IVVGFKLKHY VDEEGEKKVN YEPYLTAYDL IGAKCETRDF VVNGVTSEQL F GMCESLYV ...String:
MGSIYNYNGG CVLGMSGSNC VAIACDLRLG ANTFTTVSTK FSKIFKMNNN VYVGLSGLAT DIQTLYEILR YRVNLYEVRQ DAEMDVECF ANMLSSILYS NRFSPYFVNP IVVGFKLKHY VDEEGEKKVN YEPYLTAYDL IGAKCETRDF VVNGVTSEQL F GMCESLYV KDQDENGLFE TISQCLLSAL DRDCISGWGA EVLVLTPEKI IKKKLKARMD

UniProtKB: Proteasome subunit beta

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Macromolecule #11: 20S proteasome beta-4 subunit

MacromoleculeName: 20S proteasome beta-4 subunit / type: protein_or_peptide / ID: 11 / Number of copies: 2 / Enantiomer: LEVO / EC number: proteasome endopeptidase complex
Source (natural)Organism: Plasmodium falciparum (isolate 3D7) (eukaryote) / Strain: isolate 3D7
Molecular weightTheoretical: 22.889105 KDa
SequenceString: MDTLIGLRGN NFVVLAADTY SINSIIKLKN DDNTKFYDIH GNKCLLLGGS IGDRLQFGEF IRKNVHLYQY QNNTDMFVKS FAFFTRKNL AYYLRRNPFE VNCLIAGYDK KDGYQLYWCD YLSNMDSVNK GAHGYGAYLV SAILDKYYHE NLTVDEALDI F KLCFEELK ...String:
MDTLIGLRGN NFVVLAADTY SINSIIKLKN DDNTKFYDIH GNKCLLLGGS IGDRLQFGEF IRKNVHLYQY QNNTDMFVKS FAFFTRKNL AYYLRRNPFE VNCLIAGYDK KDGYQLYWCD YLSNMDSVNK GAHGYGAYLV SAILDKYYHE NLTVDEALDI F KLCFEELK KRFLLTQINY ELRIMYDNKV ETQYVTV

UniProtKB: Proteasome subunit beta

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Macromolecule #12: 20S proteasome beta-5 subunit

MacromoleculeName: 20S proteasome beta-5 subunit / type: protein_or_peptide / ID: 12 / Number of copies: 2 / Enantiomer: LEVO / EC number: proteasome endopeptidase complex
Source (natural)Organism: Plasmodium falciparum (isolate 3D7) (eukaryote) / Strain: isolate 3D7
Molecular weightTheoretical: 23.620646 KDa
SequenceString: TTTLAFKFKD GIIVAVDSRA SMGSFISSQN VEKIIEINKN ILGTMAGGAA DCLYWEKYLG KIIKIYELRN NEKISVRAAS TILSNILYQ YKGYGLCCGI ILSGYDHTGF NMFYVDDSGK KVEGNLFSCG SGSTYAYSIL DSAYDYNLNL DQAVELARNA I YHATFRDG ...String:
TTTLAFKFKD GIIVAVDSRA SMGSFISSQN VEKIIEINKN ILGTMAGGAA DCLYWEKYLG KIIKIYELRN NEKISVRAAS TILSNILYQ YKGYGLCCGI ILSGYDHTGF NMFYVDDSGK KVEGNLFSCG SGSTYAYSIL DSAYDYNLNL DQAVELARNA I YHATFRDG GSGGKVRVFH IHKNGYDKII EGEDVFDLHY HYTNPEQKDQ YVM

UniProtKB: Proteasome subunit beta

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Macromolecule #13: 20S proteasome beta-6 subunit

MacromoleculeName: 20S proteasome beta-6 subunit / type: protein_or_peptide / ID: 13 / Number of copies: 2 / Enantiomer: LEVO / EC number: proteasome endopeptidase complex
Source (natural)Organism: Plasmodium falciparum (isolate 3D7) (eukaryote) / Strain: isolate 3D7
Molecular weightTheoretical: 27.301203 KDa
SequenceString: MDLILYNDNL TEKKTEKENV IEHGRGFKRW YPYIDNGGTV IGLTGKDYVI LAADTRLSLS YSIYTRFCPK ISKLTDKCII GSSGMQSDI KTLHSLLQKK IQLFVLEHSH YPDIHVIARL LCVILYSRRF FPYYAFNILA GVDENNKGVL YNYDSVGSYC E ATHSCVGS ...String:
MDLILYNDNL TEKKTEKENV IEHGRGFKRW YPYIDNGGTV IGLTGKDYVI LAADTRLSLS YSIYTRFCPK ISKLTDKCII GSSGMQSDI KTLHSLLQKK IQLFVLEHSH YPDIHVIARL LCVILYSRRF FPYYAFNILA GVDENNKGVL YNYDSVGSYC E ATHSCVGS GSQLILPILD NRVEQKNQLI KNTNFNLGDD INFVKDAITS ATERDIYTGD KTLIYVIDKM GINVNTLDLK QD

UniProtKB: Proteasome subunit beta

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Macromolecule #14: 20S proteasome beta-7 subunit

MacromoleculeName: 20S proteasome beta-7 subunit / type: protein_or_peptide / ID: 14 / Number of copies: 2 / Enantiomer: LEVO / EC number: proteasome endopeptidase complex
Source (natural)Organism: Plasmodium falciparum (isolate 3D7) (eukaryote) / Strain: isolate 3D7
Molecular weightTheoretical: 30.909893 KDa
SequenceString: MTLGPVVTGT SVIAIKYKHG IMIAADRKAS YGSYAKFQNV ERIFKINNKT VMGFSGELAD AQYLHELLTR KNINNLSEKK RKEDMYTPQ HYHSYVSRVF YVRKNRIDPL FNNIIIAGIN SQKYDNNDDN VLLYTNKNND DEQNEYKNNE EYKEIHKDDL Y IGFVDMHG ...String:
MTLGPVVTGT SVIAIKYKHG IMIAADRKAS YGSYAKFQNV ERIFKINNKT VMGFSGELAD AQYLHELLTR KNINNLSEKK RKEDMYTPQ HYHSYVSRVF YVRKNRIDPL FNNIIIAGIN SQKYDNNDDN VLLYTNKNND DEQNEYKNNE EYKEIHKDDL Y IGFVDMHG TNFCDDYITT GYARYFALTL LRDHYKDNMT EEEARILINE CLRILYFRDA TSSNFIQIVK VTSKGVEYEE PY ILPCVLN SADYVYPSTL LPPAGCMW

UniProtKB: Proteasome subunit beta

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

Concentration0.7 mg/mL
BufferpH: 7.4
Component:
ConcentrationFormulaName
100.0 mMNaClSodium Chloride
5.0 mMMgCl2Magnesium Chloride
1.0 mMDTTDithiothreitol
25.0 mMTris-HClTris hydrochloride
GridModel: Quantifoil, UltrAuFoil, R1.2/1.3 / Material: GOLD / Mesh: 200 / 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 / Details: wait time 0sec blot time 2sec blot force -1.

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Electron microscopy

MicroscopeFEI TALOS ARCTICA
Specialist opticsEnergy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV
Image recordingFilm or detector model: GATAN K2 QUANTUM (4k x 4k) / Detector mode: COUNTING / Digitization - Dimensions - Width: 3838 pixel / Digitization - Dimensions - Height: 3710 pixel / Digitization - Frames/image: 1-40 / Number grids imaged: 1 / Number real images: 5200 / Average exposure time: 10.0 sec. / Average electron dose: 32.0 e/Å2
Electron beamAcceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN
Electron opticsC2 aperture diameter: 100.0 µm / Calibrated defocus max: 3.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus min: 1.0 µm / Nominal magnification: 100000
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
Experimental equipment
Model: Talos Arctica / Image courtesy: FEI Company

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Image processing

DetailsImages were gain corrected
Particle selectionNumber selected: 212749
Details: Relion autopick from 5 class averages resulting from 2000 particle picked manually
Startup modelType of model: OTHER
Details: De novo model created from 10,000 particle picked in Relion. No symmetry implied
Final reconstructionNumber classes used: 1 / Applied symmetry - Point group: C2 (2 fold cyclic) / Algorithm: BACK PROJECTION / Resolution.type: BY AUTHOR / Resolution: 3.6 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 3.0) / Number images used: 36211
Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 2.1)
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 2.1)
Final 3D classificationNumber classes: 6 / Software - Name: RELION (ver. 2.1)
FSC plot (resolution estimation)

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Atomic model buiding 1

Initial modelPDB ID:

5fmg


Chain - Source name: PDB / Chain - Initial model type: experimental model
RefinementSpace: REAL / Protocol: AB INITIO MODEL / Overall B value: 54.23
Output model

PDB-6muw:
The structure of the Plasmodium falciparum 20S proteasome.

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