3FWH

Structure of haloalkane dehalogenase mutant Dha15 (I135F/C176Y) from Rhodococcus rhodochrous


Experimental Data Snapshot

  • Method: X-RAY DIFFRACTION
  • Resolution: 1.22 Å
  • R-Value Free: 0.162 
  • R-Value Work: 0.130 
  • R-Value Observed: 0.132 

Starting Model: experimental
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This is version 1.4 of the entry. See complete history


Literature

Atomic resolution studies of haloalkane dehalogenases DhaA04, DhaA14 and DhaA15 with engineered access tunnels.

Stsiapanava, A.Dohnalek, J.Gavira, J.A.Kuty, M.Koudelakova, T.Damborsky, J.Kuta Smatanova, I.

(2010) Acta Crystallogr D Biol Crystallogr 66: 962-969

  • DOI: https://doi.org/10.1107/S0907444910027101
  • Primary Citation of Related Structures:  
    3FBW, 3FWH, 3G9X

  • PubMed Abstract: 

    The haloalkane dehalogenase DhaA from Rhodococcus rhodochrous NCIMB 13064 is a bacterial enzyme that shows catalytic activity for the hydrolytic degradation of the highly toxic industrial pollutant 1,2,3-trichloropropane (TCP). Mutagenesis focused on the access tunnels of DhaA produced protein variants with significantly improved activity towards TCP. Three mutants of DhaA named DhaA04 (C176Y), DhaA14 (I135F) and DhaA15 (C176Y + I135F) were constructed in order to study the functional relevance of the tunnels connecting the buried active site of the protein with the surrounding solvent. All three protein variants were crystallized using the sitting-drop vapour-diffusion technique. The crystals of DhaA04 belonged to the orthorhombic space group P2(1)2(1)2(1), while the crystals of DhaA14 and DhaA15 had triclinic symmetry in space group P1. The crystal structures of DhaA04, DhaA14 and DhaA15 with ligands present in the active site were solved and refined using diffraction data to 1.23, 0.95 and 1.22 A, resolution, respectively. Structural comparisons of the wild type and the three mutants suggest that the tunnels play a key role in the processes of ligand exchange between the buried active site and the surrounding solvent.


  • Organizational Affiliation

    Institute of Physical Biology, University of South Bohemia, Nove Hrady, Czech Republic.


Macromolecules
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Entity ID: 1
MoleculeChains Sequence LengthOrganismDetailsImage
Haloalkane dehalogenase299Rhodococcus sp. (in: high G+C Gram-positive bacteria)Mutation(s): 2 
Gene Names: dhaA
EC: 3.8.1.5
UniProt
Find proteins for P0A3G3 (Rhodococcus sp)
Explore P0A3G3 
Go to UniProtKB:  P0A3G3
Entity Groups  
Sequence Clusters30% Identity50% Identity70% Identity90% Identity95% Identity100% Identity
UniProt GroupP0A3G3
Sequence Annotations
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  • Reference Sequence
Experimental Data & Validation

Experimental Data

  • Method: X-RAY DIFFRACTION
  • Resolution: 1.22 Å
  • R-Value Free: 0.162 
  • R-Value Work: 0.130 
  • R-Value Observed: 0.132 
  • Space Group: P 1
Unit Cell:
Length ( Å )Angle ( ˚ )
a = 42.524α = 115.48
b = 44.37β = 98.41
c = 46.541γ = 109.6
Software Package:
Software NamePurpose
HKL-2000data collection
MOLREPphasing
REFMACrefinement
DENZOdata reduction
SCALEPACKdata scaling

Structure Validation

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Entry History 

Deposition Data

Revision History  (Full details and data files)

  • Version 1.0: 2010-02-02
    Type: Initial release
  • Version 1.1: 2011-07-13
    Changes: Version format compliance
  • Version 1.2: 2012-03-28
    Changes: Database references
  • Version 1.3: 2021-10-20
    Changes: Data collection, Database references, Derived calculations
  • Version 1.4: 2023-09-06
    Changes: Data collection, Refinement description