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- EMDB-8672: Conformational Landscape of the p28-Bound Human Proteasome Regula... -

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Basic information

Entry
Database: EMDB / ID: EMD-8672
TitleConformational Landscape of the p28-Bound Human Proteasome Regulatory Particle
Map dataThe final map of RP corrected with a B-factor of -50
Sample
  • Complex: Proteasome regulatory particle
    • Protein or peptide: x 17 types
  • Ligand: x 1 types
Keywordsp28 / 26S proteasome / regulatory particle / 19S / gankyrin / HYDROLASE
Function / homology
Function and homology information


positive regulation of inclusion body assembly / Impaired BRCA2 translocation to the nucleus / Impaired BRCA2 binding to SEM1 (DSS1) / thyrotropin-releasing hormone receptor binding / modulation by host of viral transcription / meiosis I / Hydrolases; Acting on peptide bonds (peptidases); Omega peptidases / proteasome accessory complex / integrator complex / proteasome regulatory particle ...positive regulation of inclusion body assembly / Impaired BRCA2 translocation to the nucleus / Impaired BRCA2 binding to SEM1 (DSS1) / thyrotropin-releasing hormone receptor binding / modulation by host of viral transcription / meiosis I / Hydrolases; Acting on peptide bonds (peptidases); Omega peptidases / proteasome accessory complex / integrator complex / proteasome regulatory particle / cytosolic proteasome complex / positive regulation of proteasomal protein catabolic process / proteasome regulatory particle, lid subcomplex / proteasome-activating activity / proteasome regulatory particle, base subcomplex / metal-dependent deubiquitinase activity / negative regulation of programmed cell death / protein K63-linked deubiquitination / Homologous DNA Pairing and Strand Exchange / Defective homologous recombination repair (HRR) due to BRCA1 loss of function / Defective HDR through Homologous Recombination Repair (HRR) due to PALB2 loss of BRCA1 binding function / Defective HDR through Homologous Recombination Repair (HRR) due to PALB2 loss of BRCA2/RAD51/RAD51C binding function / Resolution of D-loop Structures through Synthesis-Dependent Strand Annealing (SDSA) / Regulation of ornithine decarboxylase (ODC) / Resolution of D-loop Structures through Holliday Junction Intermediates / Cross-presentation of soluble exogenous antigens (endosomes) / K63-linked deubiquitinase activity / Somitogenesis / Impaired BRCA2 binding to RAD51 / proteasome binding / transcription factor binding / regulation of protein catabolic process / proteasome storage granule / Presynaptic phase of homologous DNA pairing and strand exchange / blastocyst development / general transcription initiation factor binding / endopeptidase activator activity / polyubiquitin modification-dependent protein binding / proteasome assembly / protein deubiquitination / positive regulation of RNA polymerase II transcription preinitiation complex assembly / mRNA export from nucleus / regulation of proteasomal protein catabolic process / enzyme regulator activity / SARS-CoV-1 targets host intracellular signalling and regulatory pathways / inclusion body / ERAD pathway / proteasome complex / Regulation of activated PAK-2p34 by proteasome mediated degradation / N-glycan trimming in the ER and Calnexin/Calreticulin cycle / Autodegradation of Cdh1 by Cdh1:APC/C / APC/C:Cdc20 mediated degradation of Securin / Asymmetric localization of PCP proteins / SCF-beta-TrCP mediated degradation of Emi1 / NIK-->noncanonical NF-kB signaling / Ubiquitin-dependent degradation of Cyclin D / AUF1 (hnRNP D0) binds and destabilizes mRNA / TNFR2 non-canonical NF-kB pathway / stem cell differentiation / Vpu mediated degradation of CD4 / Assembly of the pre-replicative complex / Degradation of DVL / Ubiquitin Mediated Degradation of Phosphorylated Cdc25A / Cdc20:Phospho-APC/C mediated degradation of Cyclin A / Dectin-1 mediated noncanonical NF-kB signaling / Hh mutants are degraded by ERAD / Degradation of AXIN / Activation of NF-kappaB in B cells / Degradation of GLI1 by the proteasome / Hedgehog ligand biogenesis / Defective CFTR causes cystic fibrosis / G2/M Checkpoints / Negative regulation of NOTCH4 signaling / GSK3B and BTRC:CUL1-mediated-degradation of NFE2L2 / Autodegradation of the E3 ubiquitin ligase COP1 / Vif-mediated degradation of APOBEC3G / double-strand break repair via homologous recombination / P-body / Regulation of RUNX3 expression and activity / Hedgehog 'on' state / Degradation of GLI2 by the proteasome / GLI3 is processed to GLI3R by the proteasome / FBXL7 down-regulates AURKA during mitotic entry and in early mitosis / MAPK6/MAPK4 signaling / APC/C:Cdh1 mediated degradation of Cdc20 and other APC/C:Cdh1 targeted proteins in late mitosis/early G1 / Degradation of beta-catenin by the destruction complex / ABC-family proteins mediated transport / Oxygen-dependent proline hydroxylation of Hypoxia-inducible Factor Alpha / HDR through Homologous Recombination (HRR) / CDK-mediated phosphorylation and removal of Cdc6 / Metalloprotease DUBs / CLEC7A (Dectin-1) signaling / SCF(Skp2)-mediated degradation of p27/p21 / double-strand break repair via nonhomologous end joining / Regulation of expression of SLITs and ROBOs / FCERI mediated NF-kB activation / Regulation of PTEN stability and activity / Interleukin-1 signaling / cytoplasmic ribonucleoprotein granule / Orc1 removal from chromatin
Similarity search - Function
: / Ubiquitin interaction motif / : / 26S proteasome regulatory subunit RPN7/PSMD6 C-terminal helix / 26S proteasome non-ATPase regulatory subunit Rpn12 / 26S proteasome regulatory subunit, C-terminal / Proteasome regulatory subunit C-terminal / DSS1/SEM1 / 26S proteasome regulatory subunit RPN5, C-terminal domain / : ...: / Ubiquitin interaction motif / : / 26S proteasome regulatory subunit RPN7/PSMD6 C-terminal helix / 26S proteasome non-ATPase regulatory subunit Rpn12 / 26S proteasome regulatory subunit, C-terminal / Proteasome regulatory subunit C-terminal / DSS1/SEM1 / 26S proteasome regulatory subunit RPN5, C-terminal domain / : / DSS1/SEM1 family / 26S proteasome regulatory subunit RPN5 C-terminal domain / PSD13 N-terminal repeats / DSS1_SEM1 / 26S proteasome regulatory subunit Rpn6, N-terminal / 6S proteasome subunit Rpn6, C-terminal helix domain / 26S proteasome regulatory subunit RPN6 N-terminal domain / 26S proteasome subunit RPN6 C-terminal helix domain / 26S Proteasome non-ATPase regulatory subunit 13 / : / 26S proteasome subunit RPN2, N-terminal domain / 26S proteasome regulatory complex, non-ATPase subcomplex, Rpn2/Psmd1 subunit / 26S proteasome regulatory subunit RPN2, C-terminal / 26S proteasome regulatory subunit RPN2 C-terminal domain / 26S Proteasome non-ATPase regulatory subunit 7/8 / : / 26S proteasome regulatory subunit 7, OB domain / : / : / PSMD12/CSN4, N-terminal / 26S proteasome regulatory subunit Rpn7/COP9 signalosome complex subunit 1 / 26S proteasome regulatory subunit Rpn7, N-terminal / 26S proteasome subunit RPN7 / 26S Proteasome non-ATPase regulatory subunit 12/COP9 signalosome complex subunit 4 / Proteasome/cyclosome repeat / Ubiquitin-interacting motif. / Proteasome/cyclosome repeat / PCI/PINT associated module / : / von Willebrand factor type A domain / HEAT repeats / Proteasomal ATPase OB C-terminal domain / Proteasomal ATPase OB C-terminal domain / CSN8/PSMD8/EIF3K / CSN8/PSMD8/EIF3K family / Rpn11/EIF3F, C-terminal / Maintenance of mitochondrial structure and function / : / motif in proteasome subunits, Int-6, Nip-1 and TRIP-15 / PCI domain / Proteasome component (PCI) domain / PCI domain profile. / Ubiquitin interacting motif / Ubiquitin-interacting motif (UIM) domain profile. / JAB1/Mov34/MPN/PAD-1 ubiquitin protease / von Willebrand factor (vWF) type A domain / VWFA domain profile. / von Willebrand factor, type A / AAA ATPase, AAA+ lid domain / AAA+ lid domain / ATPase, AAA-type, conserved site / AAA-protein family signature. / JAB/MPN domain / JAB1/MPN/MOV34 metalloenzyme domain / MPN domain / MPN domain profile. / von Willebrand factor A-like domain superfamily / ATPase family associated with various cellular activities (AAA) / ATPase, AAA-type, core / Armadillo-like helical / Tetratricopeptide-like helical domain superfamily / Armadillo-type fold / Winged helix DNA-binding domain superfamily / Winged helix-like DNA-binding domain superfamily / ATPases associated with a variety of cellular activities / AAA+ ATPase domain / Nucleic acid-binding, OB-fold / P-loop containing nucleoside triphosphate hydrolase
Similarity search - Domain/homology
26S proteasome non-ATPase regulatory subunit 11 / 26S proteasome non-ATPase regulatory subunit 12 / 26S proteasome non-ATPase regulatory subunit 14 / 26S proteasome non-ATPase regulatory subunit 3 / 26S proteasome regulatory subunit 6A / 26S proteasome regulatory subunit 7 / 26S proteasome regulatory subunit 6B / 26S proteasome non-ATPase regulatory subunit 8 / 26S proteasome non-ATPase regulatory subunit 7 / 26S proteasome non-ATPase regulatory subunit 4 ...26S proteasome non-ATPase regulatory subunit 11 / 26S proteasome non-ATPase regulatory subunit 12 / 26S proteasome non-ATPase regulatory subunit 14 / 26S proteasome non-ATPase regulatory subunit 3 / 26S proteasome regulatory subunit 6A / 26S proteasome regulatory subunit 7 / 26S proteasome regulatory subunit 6B / 26S proteasome non-ATPase regulatory subunit 8 / 26S proteasome non-ATPase regulatory subunit 7 / 26S proteasome non-ATPase regulatory subunit 4 / 26S proteasome complex subunit SEM1 / 26S proteasome regulatory subunit 4 / 26S proteasome regulatory subunit 8 / 26S proteasome regulatory subunit 10B / 26S proteasome non-ATPase regulatory subunit 6 / 26S proteasome non-ATPase regulatory subunit 1 / 26S proteasome non-ATPase regulatory subunit 13
Similarity search - Component
Biological speciesHomo sapiens (human)
Methodsingle particle reconstruction / cryo EM / Resolution: 4.5 Å
AuthorsLu Y / Wu J
CitationJournal: Mol Cell / Year: 2017
Title: Conformational Landscape of the p28-Bound Human Proteasome Regulatory Particle.
Authors: Ying Lu / Jiayi Wu / Yuanchen Dong / Shuobing Chen / Shuangwu Sun / Yong-Bei Ma / Qi Ouyang / Daniel Finley / Marc W Kirschner / Youdong Mao /
Abstract: The proteasome holoenzyme is activated by its regulatory particle (RP) consisting of two subcomplexes, the lid and the base. A key event in base assembly is the formation of a heterohexameric ring of ...The proteasome holoenzyme is activated by its regulatory particle (RP) consisting of two subcomplexes, the lid and the base. A key event in base assembly is the formation of a heterohexameric ring of AAA-ATPases, which is guided by at least four RP assembly chaperones in mammals: PAAF1, p28/gankyrin, p27/PSMD9, and S5b. Using cryogenic electron microscopy, we analyzed the non-AAA structure of the p28-bound human RP at 4.5 Å resolution and determined seven distinct conformations of the Rpn1-p28-AAA subcomplex within the p28-bound RP at subnanometer resolutions. Remarkably, the p28-bound AAA ring does not form a channel in the free RP and spontaneously samples multiple "open" and "closed" topologies at the Rpt2-Rpt6 and Rpt3-Rpt4 interfaces. Our analysis suggests that p28 assists the proteolytic core particle to select a specific conformation of the ATPase ring for RP engagement and is released in a shoehorn-like fashion in the last step of the chaperone-mediated proteasome assembly.
History
DepositionApr 12, 2017-
Header (metadata) releaseAug 23, 2017-
Map releaseAug 23, 2017-
UpdateAug 16, 2023-
Current statusAug 16, 2023Processing site: RCSB / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.0047
  • Imaged by UCSF Chimera
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  • Surface view colored by radius
  • Surface level: 0.0047
  • Imaged by UCSF Chimera
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  • Surface view with fitted model
  • Atomic models: PDB-5vgz
  • Surface level: 0.0047
  • Imaged by UCSF Chimera
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Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

Downloads & links

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Map

FileDownload / File: emd_8672.map.gz / Format: CCP4 / Size: 64 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationThe final map of RP corrected with a B-factor of -50
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
0.98 Å/pix.
x 256 pix.
= 250.88 Å
0.98 Å/pix.
x 256 pix.
= 250.88 Å
0.98 Å/pix.
x 256 pix.
= 250.88 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 0.98 Å
Density
Contour LevelBy AUTHOR: 0.004 / Movie #1: 0.0047
Minimum - Maximum-0.004617816 - 0.014717506
Average (Standard dev.)0.0002920385 (±0.0013613761)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions256256256
Spacing256256256
CellA=B=C: 250.88 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z0.980.980.98
M x/y/z256256256
origin x/y/z0.0000.0000.000
length x/y/z250.880250.880250.880
α/β/γ90.00090.00090.000
start NX/NY/NZ-34-26-36
NX/NY/NZ528549
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS256256256
D min/max/mean-0.0050.0150.000

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Supplemental data

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Sample components

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Entire : Proteasome regulatory particle

EntireName: Proteasome regulatory particle
Components
  • Complex: Proteasome regulatory particle
    • Protein or peptide: 26S proteasome regulatory subunit 7
    • Protein or peptide: 26S proteasome regulatory subunit 4
    • Protein or peptide: 26S proteasome regulatory subunit 8
    • Protein or peptide: 26S proteasome regulatory subunit 6B
    • Protein or peptide: 26S proteasome regulatory subunit 10B
    • Protein or peptide: 26S proteasome regulatory subunit 6A
    • Protein or peptide: 26S proteasome non-ATPase regulatory subunit 1
    • Protein or peptide: 26S proteasome non-ATPase regulatory subunit 3
    • Protein or peptide: 26S proteasome non-ATPase regulatory subunit 12
    • Protein or peptide: 26S proteasome non-ATPase regulatory subunit 11
    • Protein or peptide: 26S proteasome non-ATPase regulatory subunit 6
    • Protein or peptide: 26S proteasome non-ATPase regulatory subunit 7
    • Protein or peptide: 26S proteasome non-ATPase regulatory subunit 13
    • Protein or peptide: 26S proteasome non-ATPase regulatory subunit 4
    • Protein or peptide: 26S proteasome non-ATPase regulatory subunit 14
    • Protein or peptide: 26S proteasome non-ATPase regulatory subunit 8
    • Protein or peptide: 26S proteasome complex subunit SEM1
  • Ligand: ZINC ION

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Supramolecule #1: Proteasome regulatory particle

SupramoleculeName: Proteasome regulatory particle / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#17
Source (natural)Organism: Homo sapiens (human)

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Macromolecule #1: 26S proteasome regulatory subunit 7

MacromoleculeName: 26S proteasome regulatory subunit 7 / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 9.309589 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString:
APPALWDLAA DKQTLQSEQP LQVARCTKII NADSEDPKYI INVKQFAKFV VDLSDQVAPT DIEEGMRVGV DRNKYQIHIP LPP

UniProtKB: 26S proteasome regulatory subunit 7

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Macromolecule #2: 26S proteasome regulatory subunit 4

MacromoleculeName: 26S proteasome regulatory subunit 4 / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 7.969963 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString:
EEERSKVDDL RGTPMSVGTL EEIIDDNHAI VSTSVGSEHY VSILSFVDKD LLEPGCSVLL NHKVHAVIGV LMD

UniProtKB: 26S proteasome regulatory subunit 4

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Macromolecule #3: 26S proteasome regulatory subunit 8

MacromoleculeName: 26S proteasome regulatory subunit 8 / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 13.603661 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString:
LEEGKAGSGL RQYYLSKIEE LQLIVNDKSQ NLRRLQAQRN ELNAKVRLLR EELQLLQEQG SYVGEVVRAM DKKKVLVKVH PEGKFVVDV DKNIDINDVT PNCRVALRND SYTLHKILP

UniProtKB: 26S proteasome regulatory subunit 8

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Macromolecule #4: 26S proteasome regulatory subunit 6B

MacromoleculeName: 26S proteasome regulatory subunit 6B / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 12.341005 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString:
DLYSRYKKLQ QELEFLEVQE EYIKDEQKNL KKEFLHAQEE VKRIQSIPLV IGQFLEAVDQ NTAIVGSTTG SNYYVRILST IDRELLKPN ASVALHKHSN ALVDVLPP

UniProtKB: 26S proteasome regulatory subunit 6B

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Macromolecule #5: 26S proteasome regulatory subunit 10B

MacromoleculeName: 26S proteasome regulatory subunit 10B / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 12.197259 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString:
DYRKKLLEHK EIDGRLKELR EQLKELTKQY EKSENDLKAL QSVGQIVGEV LKQLTEEKFI VKATNGPRYV VGCRRQLDKS KLKPGTRVA LDMTTLTIMR YLPRE

UniProtKB: 26S proteasome regulatory subunit 10B

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Macromolecule #6: 26S proteasome regulatory subunit 6A

MacromoleculeName: 26S proteasome regulatory subunit 6A / type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 12.942888 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString:
KIMKSEVLRV THELQAMKDK IKENSEKIKV NKTLPYLVSN VIELLDVDPN DQEEDGANID LDSQRKGKCA VIKTSTRQTY FLPVIGLVD AEKLKPGDLV GVNKDSYLIL ETLPTE

UniProtKB: 26S proteasome regulatory subunit 6A

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Macromolecule #7: 26S proteasome non-ATPase regulatory subunit 1

MacromoleculeName: 26S proteasome non-ATPase regulatory subunit 1 / type: protein_or_peptide / ID: 7 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 103.755023 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: MITSAAGIIS LLDEDEPQLK EFALHKLNAV VNDFWAEISE SVDKIEVLYE DEGFRSRQFA ALVASKVFYH LGAFEESLNY ALGAGDLFN VNDNSEYVET IIAKCIDHYT KQCVENADLP EGEKKPIDQR LEGIVNKMFQ RCLDDHKYKQ AIGIALETRR L DVFEKTIL ...String:
MITSAAGIIS LLDEDEPQLK EFALHKLNAV VNDFWAEISE SVDKIEVLYE DEGFRSRQFA ALVASKVFYH LGAFEESLNY ALGAGDLFN VNDNSEYVET IIAKCIDHYT KQCVENADLP EGEKKPIDQR LEGIVNKMFQ RCLDDHKYKQ AIGIALETRR L DVFEKTIL ESNDVPGMLA YSLKLCMSLM QNKQFRNKVL RVLVKIYMNL EKPDFINVCQ CLIFLDDPQA VSDILEKLVK ED NLLMAYQ ICFDLYESAS QQFLSSVIQN LRTVGTPIAS VPGSTNTGTV PGSEKDSDSM ETEEKTSSAF VGKTPEASPE PKD QTLKMI KILSGEMAIE LHLQFLIRNN NTDLMILKNT KDAVRNSVCH TATVIANSFM HCGTTSDQFL RDNLEWLARA TNWA KFTAT ASLGVIHKGH EKEALQLMAT YLPKDTSPGS AYQEGGGLYA LGLIHANHGG DIIDYLLNQL KNASNDIVRH GGSLG LGLA AMGTARQDVY DLLKTNLYQD DAVTGEAAGL ALGLVMLGSK NAQAIEDMVG YAQETQHEKI LRGLAVGIAL VMYGRM EEA DALIESLCRD KDPILRRSGM YTVAMAYCGS GNNKAIRRLL HVAVSDVNDD VRRAAVESLG FILFRTPEQC PSVVSLL SE SYNPHVRYGA AMALGICCAG TGNKEAINLL EPMTNDPVNY VRQGALIASA LIMIQQTEIT CPKVNQFRQL YSKVINDK H DDVMAKFGAI LAQGILDAGG HNVTISLQSR TGHTHMPSVV GVLVFTQFWF WFPLSHFLSL AYTPTCVIGL NKDLKMPKV QYKSNCKPST FAYPAPLEVP KEKEKEKVST AVLSITAKAK KKEKEKEKKE EEKMEVDEAE KKEEKEKKKE PEPNFQLLDN PARVMPAQL KVLTMPETCR YQPFKPLSIG GIIILKDTSE DIEELVEPVA AHGPKI

UniProtKB: 26S proteasome non-ATPase regulatory subunit 1

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Macromolecule #8: 26S proteasome non-ATPase regulatory subunit 3

MacromoleculeName: 26S proteasome non-ATPase regulatory subunit 3 / type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 55.703727 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: PPGGGEQEPP PPPAPQDVEM KEEAATGGGS TGEADGKTAA AAAEHSQREL DTVTLEDIKE HVKQLEKAVS GKEPRFVLRA LRMLPSTSR RLNHYVLYKA VQGFFTSNNA TRDFLLPFLE EPMDTEADLQ FRPRTGKAAS TPLLPEVEAY LQLLVVIFMM N SKRYKEAQ ...String:
PPGGGEQEPP PPPAPQDVEM KEEAATGGGS TGEADGKTAA AAAEHSQREL DTVTLEDIKE HVKQLEKAVS GKEPRFVLRA LRMLPSTSR RLNHYVLYKA VQGFFTSNNA TRDFLLPFLE EPMDTEADLQ FRPRTGKAAS TPLLPEVEAY LQLLVVIFMM N SKRYKEAQ KISDDLMQKI STQNRRALDL VAAKCYYYHA RVYEFLDKLD VVRSFLHARL RTATLRHDAD GQATLLNLLL RN YLHYSLY DQAEKLVSKS VFPEQANNNE WARYLYYTGR IKAIQLEYSE ARRTMTNALR KAPQHTAVGF KQTVHKLLIV VEL LLGEIP DRLQFRQPSL KRSLMPYFLL TQAVRTGNLA KFNQVLDQFG EKFQADGTYT LIIRLRHNVI KTGVRMISLS YSRI SLADI AQKLQLDSPE DAEFIVAKAI RDGVIEASIN HEKGYVQSKE MIDIYSTREP QLAFHQRISF CLDIHNMSVK AMRFP PKSY NKDL

UniProtKB: 26S proteasome non-ATPase regulatory subunit 3

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Macromolecule #9: 26S proteasome non-ATPase regulatory subunit 12

MacromoleculeName: 26S proteasome non-ATPase regulatory subunit 12 / type: protein_or_peptide / ID: 9 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 52.979359 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: MADGGSERAD GRIVKMEVDY SATVDQRLPE CAKLAKEGRL QEVIETLLSL EKQTRTASDM VSTSRILVAV VKMCYEAKEW DLLNENIML LSKRRSQLKQ AVAKMVQQCC TYVEEITDLP IKLRLIDTLR MVTEGKIYVE IERARLTKTL ATIKEQNGDV K EAASILQE ...String:
MADGGSERAD GRIVKMEVDY SATVDQRLPE CAKLAKEGRL QEVIETLLSL EKQTRTASDM VSTSRILVAV VKMCYEAKEW DLLNENIML LSKRRSQLKQ AVAKMVQQCC TYVEEITDLP IKLRLIDTLR MVTEGKIYVE IERARLTKTL ATIKEQNGDV K EAASILQE LQVETYGSME KKERVEFILE QMRLCLAVKD YIRTQIISKK INTKFFQEEN TEKLKLKYYN LMIQLDQHEG SY LSICKHY RAIYDTPCIQ AESEKWQQAL KSVVLYVILA PFDNEQSDLV HRISGDKKLE EIPKYKDLLK LFTTMELMRW STL VEDYGM ELRKGSLESP ATDVFGSTEE GEKRWKDLKN RVVEHNIRIM AKYYTRITMK RMAQLLDLSV DESEAFLSNL VVNK TIFAK VDRLAGIINF QRPKDPNNLL NDWSQKLNSL MSLVNKTTHL IAKEEMIHNL Q

UniProtKB: 26S proteasome non-ATPase regulatory subunit 12

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Macromolecule #10: 26S proteasome non-ATPase regulatory subunit 11

MacromoleculeName: 26S proteasome non-ATPase regulatory subunit 11 / type: protein_or_peptide / ID: 10 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 43.427055 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: NDEEAVQVKE QSILELGSLL AKTGQAAELG GLLKYVRPFL NSISKAKAAR LVRSLLDLFL DMEAATGQEV ELCLECIEWA KSEKRTFLR QALEARLVSL YFDTKRYQEA LHLGSQLLRE LKKMDDKALL VEVQLLESKT YHALSNLPKA RAALTSARTT A NAIYCPPK ...String:
NDEEAVQVKE QSILELGSLL AKTGQAAELG GLLKYVRPFL NSISKAKAAR LVRSLLDLFL DMEAATGQEV ELCLECIEWA KSEKRTFLR QALEARLVSL YFDTKRYQEA LHLGSQLLRE LKKMDDKALL VEVQLLESKT YHALSNLPKA RAALTSARTT A NAIYCPPK LQATLDMQSG IIHAAEEKDW KTAYSYFYEA FEGYDSIDSP KAITSLKYML LCKIMLNTPE DVQALVSGKL AL RYAGRQT EALKCVAQAS KNRSLADFEK ALTDYRAELR DDPIISTHLA KLYDNLLEQN LIRVIEPFSR VQIEHISSLI KLS KADVER KLSQMILDKK FHGILDQGEG VLIIFDEPPV DKTYEAALET IQNMSKVVDS LYNKAKKLT

UniProtKB: 26S proteasome non-ATPase regulatory subunit 11

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Macromolecule #11: 26S proteasome non-ATPase regulatory subunit 6

MacromoleculeName: 26S proteasome non-ATPase regulatory subunit 6 / type: protein_or_peptide / ID: 11 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 44.336906 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: PKNPDLRIAQ LRFLLSLPEH RGDAAVRDEL MAAVRDNNMA PYYEALCKSL DWQIDVDLLN KMKKANEDEL KRLDEELEDA EKNLGESEI RDAMMAKAEY LCRIGDKEGA LTAFRKTYDK TVALGHRLDI VFYLLRIGLF YMDNDLITRN TEKAKSLIEE G GDWDRRNR ...String:
PKNPDLRIAQ LRFLLSLPEH RGDAAVRDEL MAAVRDNNMA PYYEALCKSL DWQIDVDLLN KMKKANEDEL KRLDEELEDA EKNLGESEI RDAMMAKAEY LCRIGDKEGA LTAFRKTYDK TVALGHRLDI VFYLLRIGLF YMDNDLITRN TEKAKSLIEE G GDWDRRNR LKVYQGLYCV AIRDFKQAAE LFLDTVSTFT SYELMDYKTF VTYTVYVSMI ALERPDLREK VIKGAEILEV LH SLPAVRQ YLFSLYECRY SVFFQSLAVV EQEMKKDWLF APHYRYYVRE MRIHAYSQLL ESYRSLTLGY MAEAFGVGVE FID QELSRF IAAGRLHCKI DKVNEIVETN RPDSKNWQYQ ETIKKGDLLL NRVQKLSRVI NM

UniProtKB: 26S proteasome non-ATPase regulatory subunit 6

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Macromolecule #12: 26S proteasome non-ATPase regulatory subunit 7

MacromoleculeName: 26S proteasome non-ATPase regulatory subunit 7 / type: protein_or_peptide / ID: 12 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 32.382094 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: AVQKVVVHPL VLLSVVDHFN RIGKVGNQKR VVGVLLGSWQ KKVLDVSNSF AVPFDEDDKD DSVWFLDHDY LENMYGMFKK VNARERIVG WYHTGPKLHK NDIAINELMK RYCPNSVLVI IDVKPKDLGL PTEAYISVEE VHDDGTPTSK TFEHVTSEIG A EEAEEVGV ...String:
AVQKVVVHPL VLLSVVDHFN RIGKVGNQKR VVGVLLGSWQ KKVLDVSNSF AVPFDEDDKD DSVWFLDHDY LENMYGMFKK VNARERIVG WYHTGPKLHK NDIAINELMK RYCPNSVLVI IDVKPKDLGL PTEAYISVEE VHDDGTPTSK TFEHVTSEIG A EEAEEVGV EHLLRDIKDT TVGTLSQRIT NQVHGLKGLN SKLLDIRSYL EKVATGKLPI NHQIIYQLQD VFNLLPDVSL QE FVKAFYL KTNDQMVVVY LASLIRSVVA LHNLINNKIA NRDAEKKEG

UniProtKB: 26S proteasome non-ATPase regulatory subunit 7

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Macromolecule #13: 26S proteasome non-ATPase regulatory subunit 13

MacromoleculeName: 26S proteasome non-ATPase regulatory subunit 13 / type: protein_or_peptide / ID: 13 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 42.734988 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: DVPGFLQQSQ NSGPGQPAVW HRLEELYTKK LWHQLTLQVL DFVQDPCFAQ GDGLIKLYEN FISEFEHRVN PLSLVEIILH VVRQMTDPN VALTFLEKTR EKVKSSDEAV ILCKTAIGAL KLNIGDLQVT KETIEDVEEM LNNLPGVTSV HSRFYDLSSK Y YQTIGNHA ...String:
DVPGFLQQSQ NSGPGQPAVW HRLEELYTKK LWHQLTLQVL DFVQDPCFAQ GDGLIKLYEN FISEFEHRVN PLSLVEIILH VVRQMTDPN VALTFLEKTR EKVKSSDEAV ILCKTAIGAL KLNIGDLQVT KETIEDVEEM LNNLPGVTSV HSRFYDLSSK Y YQTIGNHA SYYKDALRFL GCVDIKDLPV SEQQERAFTL GLAGLLGEGV FNFGELLMHP VLESLRNTDR QWLIDTLYAF NS GNVERFQ TLKTAWGQQP DLAANEAQLL RKIQLLCLME MTFTRPANHR QLTFEEIAKS AKITVNEVEL LVMKALSVGL VKG SIDEVD KRVHMTWVQP RVLDLQQIKG MKDRLEFWCT DVKSMEMLVE HQAHDILT

UniProtKB: 26S proteasome non-ATPase regulatory subunit 13

+
Macromolecule #14: 26S proteasome non-ATPase regulatory subunit 4

MacromoleculeName: 26S proteasome non-ATPase regulatory subunit 4 / type: protein_or_peptide / ID: 14 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 20.866023 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString:
MVLESTMVCV DNSEYMRNGD FLPTRLQAQQ DAVNIVCHSK TRSNPENNVG LITLANDCEV LTTLTPDTGR ILSKLHTVQP KGKITFCTG IRVAHLALKH RQGKNHKMRI IAFVGSPVED NEKDLVKLAK RLKKEKVNVD IINFGEEEVN TEKLTAFVNT L NGKDGTGS HLVTVPPGPS LADALISSPI LAG

UniProtKB: 26S proteasome non-ATPase regulatory subunit 4

+
Macromolecule #15: 26S proteasome non-ATPase regulatory subunit 14

MacromoleculeName: 26S proteasome non-ATPase regulatory subunit 14 / type: protein_or_peptide / ID: 15 / Number of copies: 1 / Enantiomer: LEVO
EC number: Hydrolases; Acting on peptide bonds (peptidases); Omega peptidases
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 32.329355 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: AVDTAEQVYI SSLALLKMLK HGRAGVPMEV MGLMLGEFVD DYTVRVIDVF AMPQSGTGVS VEAVDPVFQA KMLDMLKQTG RPEMVVGWY HSHPGFGCWL SGVDINTQQS FEALSERAVA VVVDPIQSVK GKVVIDAFRL INANMMVLGH EPRQTTSNLG H LNKPSIQA ...String:
AVDTAEQVYI SSLALLKMLK HGRAGVPMEV MGLMLGEFVD DYTVRVIDVF AMPQSGTGVS VEAVDPVFQA KMLDMLKQTG RPEMVVGWY HSHPGFGCWL SGVDINTQQS FEALSERAVA VVVDPIQSVK GKVVIDAFRL INANMMVLGH EPRQTTSNLG H LNKPSIQA LIHGLNRHYY SITINYRKNE LEQKMLLNLH KKSWMEGLTL QDYSEHCKHN ESVVKEMLEL AKNYNKAVEE ED KMTPEQL AIKNVGKQDP KRHLEEHVDV LMTSNIVQCL AAMLDTVVFK

UniProtKB: 26S proteasome non-ATPase regulatory subunit 14

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Macromolecule #16: 26S proteasome non-ATPase regulatory subunit 8

MacromoleculeName: 26S proteasome non-ATPase regulatory subunit 8 / type: protein_or_peptide / ID: 16 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 30.039699 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: MYEQLKGEWN RKSPNLSKCG EELGRLKLVL LELNFLPTTG TKLTKQQLIL ARDILEIGAQ WSILRKDIPS FERYMAQLKC YYFDYKEQL PESAYMHQLL GLNLLFLLSQ NRVAEFHTEL ERLPAKDIQT NVYIKHPVSL EQYLMEGSYN KVFLAKGNIP A ESYTFFID ...String:
MYEQLKGEWN RKSPNLSKCG EELGRLKLVL LELNFLPTTG TKLTKQQLIL ARDILEIGAQ WSILRKDIPS FERYMAQLKC YYFDYKEQL PESAYMHQLL GLNLLFLLSQ NRVAEFHTEL ERLPAKDIQT NVYIKHPVSL EQYLMEGSYN KVFLAKGNIP A ESYTFFID ILLDTIRDEI AGCIEKAYEK ILFTEATRIL FFNTPKKMTD YAKKRGWVLG PNNYYSFASQ QQKPEDTTIP ST ELAKQVI EYARQLEMIV

UniProtKB: 26S proteasome non-ATPase regulatory subunit 8

+
Macromolecule #17: 26S proteasome complex subunit SEM1

MacromoleculeName: 26S proteasome complex subunit SEM1 / type: protein_or_peptide / ID: 17 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 8.284611 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString:
MSEKKQPVDL GLLEEDDEFE EFPAEDWAGL DEDEDAHVWE DNWDDDNVED DFSNQLRAEL EKHGYKMETS

UniProtKB: 26S proteasome complex subunit SEM1

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Macromolecule #18: ZINC ION

MacromoleculeName: ZINC ION / type: ligand / ID: 18 / Number of copies: 1 / Formula: ZN
Molecular weightTheoretical: 65.409 Da

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

BufferpH: 7.5
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeFEI TECNAI ARCTICA
Image recordingFilm or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 50.0 e/Å2
Electron beamAcceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD
Experimental equipment
Model: Talos Arctica / Image courtesy: FEI Company

-
Image processing

Startup modelType of model: INSILICO MODEL
Final reconstructionResolution.type: BY AUTHOR / Resolution: 4.5 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 117471
Initial angle assignmentType: ANGULAR RECONSTITUTION
Final angle assignmentType: PROJECTION MATCHING

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