2NVU

Structure of APPBP1-UBA3~NEDD8-NEDD8-MgATP-Ubc12(C111A), a trapped ubiquitin-like protein activation complex


Experimental Data Snapshot

  • Method: X-RAY DIFFRACTION
  • Resolution: 2.80 Å
  • R-Value Free: 0.274 
  • R-Value Work: 0.241 
  • R-Value Observed: 0.241 

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This is version 1.4 of the entry. See complete history


Literature

Basis for a ubiquitin-like protein thioester switch toggling E1-E2 affinity.

Huang, D.T.Hunt, H.W.Zhuang, M.Ohi, M.D.Holton, J.M.Schulman, B.A.

(2007) Nature 445: 394-398

  • DOI: https://doi.org/10.1038/nature05490
  • Primary Citation of Related Structures:  
    2NVU

  • PubMed Abstract: 

    Ubiquitin-like proteins (UBLs) are conjugated by dynamic E1-E2-E3 enzyme cascades. E1 enzymes activate UBLs by catalysing UBL carboxy-terminal adenylation, forming a covalent E1 throught UBL thioester intermediate, and generating a thioester-linked E2 throught UBL product, which must be released for subsequent reactions. Here we report the structural analysis of a trapped UBL activation complex for the human NEDD8 pathway, containing NEDD8's heterodimeric E1 (APPBP1-UBA3), two NEDD8s (one thioester-linked to E1, one noncovalently associated for adenylation), a catalytically inactive E2 (Ubc12), and MgATP. The results suggest that a thioester switch toggles E1-E2 affinities. Two E2 binding sites depend on NEDD8 being thioester-linked to E1. One is unmasked by a striking E1 conformational change. The other comes directly from the thioester-bound NEDD8. After NEDD8 transfer to E2, reversion to an alternate E1 conformation would facilitate release of the E2 throught NEDD8 thioester product. Thus, transferring the UBL's thioester linkage between successive conjugation enzymes can induce conformational changes and alter interaction networks to drive consecutive steps in UBL cascades.


  • Organizational Affiliation

    Howard Hughes Medical Institute, St Jude Children's Research Hospital, Memphis, Tennessee 38105, USA.


Macromolecules
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Entity ID: 1
MoleculeChains Sequence LengthOrganismDetailsImage
NEDD8-activating enzyme E1 regulatory subunit536Homo sapiensMutation(s): 0 
Gene Names: APPBP1
UniProt & NIH Common Fund Data Resources
Find proteins for Q13564 (Homo sapiens)
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Go to UniProtKB:  Q13564
PHAROS:  Q13564
GTEx:  ENSG00000159593 
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UniProt GroupQ13564
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  • Reference Sequence
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Entity ID: 2
MoleculeChains Sequence LengthOrganismDetailsImage
Maltose binding protein/NEDD8-activating enzyme E1 catalytic subunit chimera805Homo sapiensMutation(s): 0 
Gene Names: UBE1CUBA3
EC: 6.3.2
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Find proteins for Q8TBC4 (Homo sapiens)
Explore Q8TBC4 
Go to UniProtKB:  Q8TBC4
PHAROS:  Q8TBC4
GTEx:  ENSG00000144744 
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UniProt GroupQ8TBC4
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  • Reference Sequence
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Entity ID: 3
MoleculeChains Sequence LengthOrganismDetailsImage
NEDD8-conjugating enzyme Ubc12180Homo sapiensMutation(s): 1 
Gene Names: UBE2MUBC12
EC: 6.3.2
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Find proteins for P61081 (Homo sapiens)
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PHAROS:  P61081
GTEx:  ENSG00000130725 
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  • Reference Sequence
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Entity ID: 4
MoleculeChains Sequence LengthOrganismDetailsImage
NEDD8D [auth I],
E [auth J]
81Homo sapiensMutation(s): 0 
Gene Names: NEDD8
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Find proteins for Q15843 (Homo sapiens)
Explore Q15843 
Go to UniProtKB:  Q15843
PHAROS:  Q15843
GTEx:  ENSG00000129559 
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UniProt GroupQ15843
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  • Reference Sequence
Experimental Data & Validation

Experimental Data

  • Method: X-RAY DIFFRACTION
  • Resolution: 2.80 Å
  • R-Value Free: 0.274 
  • R-Value Work: 0.241 
  • R-Value Observed: 0.241 
  • Space Group: P 32 2 1
Unit Cell:
Length ( Å )Angle ( ˚ )
a = 156.494α = 90
b = 156.494β = 90
c = 190.485γ = 120
Software Package:
Software NamePurpose
CNSrefinement
PDB_EXTRACTdata extraction
HKL-2000data reduction
SCALEPACKdata scaling
PHASERphasing

Structure Validation

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Ligand Structure Quality Assessment 


Entry History 

Deposition Data

Revision History  (Full details and data files)

  • Version 1.0: 2007-01-30
    Type: Initial release
  • Version 1.1: 2008-05-01
    Changes: Version format compliance
  • Version 1.2: 2011-07-13
    Changes: Version format compliance
  • Version 1.3: 2021-10-20
    Changes: Database references, Derived calculations
  • Version 1.4: 2023-08-30
    Changes: Data collection, Refinement description