2YFI

Crystal Structure of Biphenyl dioxygenase variant RR41 (BPDO-RR41)


Experimental Data Snapshot

  • Method: X-RAY DIFFRACTION
  • Resolution: 2.15 Å
  • R-Value Free: 0.232 
  • R-Value Work: 0.200 
  • R-Value Observed: 0.202 

wwPDB Validation   3D Report Full Report


This is version 1.2 of the entry. See complete history


Literature

Retuning Rieske-Type Oxygenases to Expand Substrate Range.

Mohammadi, M.Viger, J.Kumar, P.Barriault, D.Bolin, J.T.Sylvestre, M.

(2011) J Biol Chem 286: 27612

  • DOI: https://doi.org/10.1074/jbc.M111.255174
  • Primary Citation of Related Structures:  
    2YFI, 2YFJ

  • PubMed Abstract: 

    Rieske-type oxygenases are promising biocatalysts for the destruction of persistent pollutants or for the synthesis of fine chemicals. In this work, we explored pathways through which Rieske-type oxygenases evolve to expand their substrate range. BphAE(p4), a variant biphenyl dioxygenase generated from Burkholderia xenovorans LB400 BphAE(LB400) by the double substitution T335A/F336M, and BphAE(RR41), obtained by changing Asn(338), Ile(341), and Leu(409) of BphAE(p4) to Gln(338), Val(341), and Phe(409), metabolize dibenzofuran two and three times faster than BphAE(LB400), respectively. Steady-state kinetic measurements of single- and multiple-substitution mutants of BphAE(LB400) showed that the single T335A and the double N338Q/L409F substitutions contribute significantly to enhanced catalytic activity toward dibenzofuran. Analysis of crystal structures showed that the T335A substitution relieves constraints on a segment lining the catalytic cavity, allowing a significant displacement in response to dibenzofuran binding. The combined N338Q/L409F substitutions alter substrate-induced conformational changes of protein groups involved in subunit assembly and in the chemical steps of the reaction. This suggests a responsive induced fit mechanism that retunes the alignment of protein atoms involved in the chemical steps of the reaction. These enzymes can thus expand their substrate range through mutations that alter the constraints or plasticity of the catalytic cavity to accommodate new substrates or that alter the induced fit mechanism required to achieve proper alignment of reaction-critical atoms or groups.


  • Organizational Affiliation

    Institut National de la Recherche Scientifique-Institut Armand-Frappier, Laval, Quebec H7V 1B7, Canada.


Macromolecules
Find similar proteins by:  (by identity cutoff)  |  3D Structure
Entity ID: 1
MoleculeChains Sequence LengthOrganismDetailsImage
BIPHENYL DIOXYGENASE SUBUNIT ALPHA
A, C, E, G, I
A, C, E, G, I, K
459Paraburkholderia xenovorans LB400Mutation(s): 5 
EC: 1.14.12.18
UniProt
Find proteins for P37333 (Paraburkholderia xenovorans (strain LB400))
Explore P37333 
Go to UniProtKB:  P37333
Entity Groups  
Sequence Clusters30% Identity50% Identity70% Identity90% Identity95% Identity100% Identity
UniProt GroupP37333
Sequence Annotations
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  • Reference Sequence
Find similar proteins by:  (by identity cutoff)  |  3D Structure
Entity ID: 2
MoleculeChains Sequence LengthOrganismDetailsImage
BIPHENYL DIOXYGENASE SUBUNIT BETA
B, D, F, H, J
B, D, F, H, J, L
188Paraburkholderia xenovorans LB400Mutation(s): 0 
EC: 1.14.12.18
UniProt
Find proteins for P37334 (Paraburkholderia xenovorans (strain LB400))
Explore P37334 
Go to UniProtKB:  P37334
Entity Groups  
Sequence Clusters30% Identity50% Identity70% Identity90% Identity95% Identity100% Identity
UniProt GroupP37334
Sequence Annotations
Expand
  • Reference Sequence
Small Molecules
Ligands 2 Unique
IDChains Name / Formula / InChI Key2D Diagram3D Interactions
FES
Query on FES

Download Ideal Coordinates CCD File 
M [auth A]
O [auth C]
Q [auth E]
S [auth G]
U [auth I]
M [auth A],
O [auth C],
Q [auth E],
S [auth G],
U [auth I],
W [auth K]
FE2/S2 (INORGANIC) CLUSTER
Fe2 S2
NIXDOXVAJZFRNF-UHFFFAOYSA-N
FE2
Query on FE2

Download Ideal Coordinates CCD File 
N [auth A]
P [auth C]
R [auth E]
T [auth G]
V [auth I]
N [auth A],
P [auth C],
R [auth E],
T [auth G],
V [auth I],
X [auth K]
FE (II) ION
Fe
CWYNVVGOOAEACU-UHFFFAOYSA-N
Experimental Data & Validation

Experimental Data

  • Method: X-RAY DIFFRACTION
  • Resolution: 2.15 Å
  • R-Value Free: 0.232 
  • R-Value Work: 0.200 
  • R-Value Observed: 0.202 
  • Space Group: P 1 21 1
Unit Cell:
Length ( Å )Angle ( ˚ )
a = 86.969α = 90
b = 277.808β = 117.61
c = 92.933γ = 90
Software Package:
Software NamePurpose
REFMACrefinement
HKL-2000data reduction
SCALEPACKdata scaling
MOLREPphasing

Structure Validation

View Full Validation Report



Entry History 

Deposition Data

Revision History  (Full details and data files)

  • Version 1.0: 2011-06-08
    Type: Initial release
  • Version 1.1: 2011-08-10
    Changes: Database references, Version format compliance
  • Version 1.2: 2023-12-20
    Changes: Data collection, Database references, Derived calculations, Other, Refinement description