3A2Z

E. coli Gsp amidase Cys59 sulfenic acid


Experimental Data Snapshot

  • Method: X-RAY DIFFRACTION
  • Resolution: 1.50 Å
  • R-Value Free: 0.204 
  • R-Value Work: 0.180 

wwPDB Validation   3D Report Full Report


This is version 1.4 of the entry. See complete history


Literature

Protein S-thiolation by Glutathionylspermidine (Gsp): the role of Escherichia coli Gsp synthetASE/amidase in redox regulation

Chiang, B.-Y.Chen, T.-C.Pai, C.-H.Chou, C.-C.Chen, H.-H.Ko, T.-P.Hsu, W.-H.Chang, C.-Y.Wu, W.-F.Wang, A.H.-J.Lin, C.-H.

(2010) J Biol Chem 285: 25345-25353

  • DOI: https://doi.org/10.1074/jbc.M110.133363
  • Primary Citation of Related Structures:  
    3A2Z, 3A30

  • PubMed Abstract: 

    Certain bacteria synthesize glutathionylspermidine (Gsp), from GSH and spermidine. Escherichia coli Gsp synthetase/amidase (GspSA) catalyzes both the synthesis and hydrolysis of Gsp. Prior to the work reported herein, the physiological role(s) of Gsp or how the two opposing GspSA activities are regulated had not been elucidated. We report that Gsp-modified proteins from E. coli contain mixed disulfides of Gsp and protein thiols, representing a new type of post-translational modification formerly undocumented. The level of these proteins is increased by oxidative stress. We attribute the accumulation of such proteins to the selective inactivation of GspSA amidase activity. X-ray crystallography and a chemical modification study indicated that the catalytic cysteine thiol of the GspSA amidase domain is transiently inactivated by H(2)O(2) oxidation to sulfenic acid, which is stabilized by a very short hydrogen bond with a water molecule. We propose a set of reactions that explains how the levels of Gsp and Gsp S-thiolated proteins are modulated in response to oxidative stress. The hypersensitivities of GspSA and GspSA/glutaredoxin null mutants to H(2)O(2) support the idea that GspSA and glutaredoxin act synergistically to regulate the redox environment of E. coli.


  • Organizational Affiliation

    Institute of Biological Chemistry, Facilities for Proteomics Research, Academia Sinica, 128 Academia Road, Section 2, Taipei 11529, Taiwan.


Macromolecules
Find similar proteins by:  (by identity cutoff)  |  3D Structure
Entity ID: 1
MoleculeChains Sequence LengthOrganismDetailsImage
Bifunctional glutathionylspermidine synthetase/amidase197Escherichia coli K-12Mutation(s): 0 
Gene Names: gspb2988JW2956
EC: 6.3.1.8 (PDB Primary Data), 3.5.1.78 (PDB Primary Data)
UniProt
Find proteins for P0AES0 (Escherichia coli (strain K12))
Explore P0AES0 
Go to UniProtKB:  P0AES0
Entity Groups  
Sequence Clusters30% Identity50% Identity70% Identity90% Identity95% Identity100% Identity
UniProt GroupP0AES0
Sequence Annotations
Expand
  • Reference Sequence
Experimental Data & Validation

Experimental Data

  • Method: X-RAY DIFFRACTION
  • Resolution: 1.50 Å
  • R-Value Free: 0.204 
  • R-Value Work: 0.180 
  • Space Group: P 64 2 2
Unit Cell:
Length ( Å )Angle ( ˚ )
a = 83.902α = 90
b = 83.902β = 90
c = 106.531γ = 120
Software Package:
Software NamePurpose
Blu-Icedata collection
CNSrefinement
HKL-2000data reduction
HKL-2000data scaling
CNSphasing

Structure Validation

View Full Validation Report



Entry History 

Deposition Data

Revision History  (Full details and data files)

  • Version 1.0: 2010-05-19
    Type: Initial release
  • Version 1.1: 2011-07-13
    Changes: Version format compliance
  • Version 1.2: 2017-10-11
    Changes: Refinement description
  • Version 1.3: 2021-03-10
    Changes: Advisory, Derived calculations
  • Version 1.4: 2023-11-01
    Changes: Data collection, Database references, Refinement description