3KVZ

Structural basis of the activity and substrate specificity of the fluoroacetyl-CoA thiesterase FlK - wild type FlK in complex with FAcCPan


Experimental Data Snapshot

  • Method: X-RAY DIFFRACTION
  • Resolution: 2.10 Å
  • R-Value Free: 0.282 
  • R-Value Work: 0.204 
  • R-Value Observed: 0.208 

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This is version 1.3 of the entry. See complete history


Literature

Structural basis for the activity and substrate specificity of fluoroacetyl-CoA thioesterase FlK.

Dias, M.V.Huang, F.Chirgadze, D.Y.Tosin, M.Spiteller, D.Dry, E.F.Leadlay, P.F.Spencer, J.B.Blundell, T.L.

(2010) J Biol Chem 285: 22495-22504

  • DOI: https://doi.org/10.1074/jbc.M110.107177
  • Primary Citation of Related Structures:  
    3KUV, 3KUW, 3KV7, 3KV8, 3KVI, 3KVU, 3KVZ, 3KW1, 3KX7, 3KX8

  • PubMed Abstract: 

    The thioesterase FlK from the fluoroacetate-producing Streptomyces cattleya catalyzes the hydrolysis of fluoroacetyl-coenzyme A. This provides an effective self-defense mechanism, preventing any fluoroacetyl-coenzyme A formed from being further metabolized to 4-hydroxy-trans-aconitate, a lethal inhibitor of the tricarboxylic acid cycle. Remarkably, FlK does not accept acetyl-coenzyme A as a substrate. Crystal structure analysis shows that FlK forms a dimer, in which each subunit adopts a hot dog fold as observed for type II thioesterases. Unlike other type II thioesterases, which invariably utilize either an aspartate or a glutamate as catalytic base, we show by site-directed mutagenesis and crystallography that FlK employs a catalytic triad composed of Thr(42), His(76), and a water molecule, analogous to the Ser/Cys-His-acid triad of type I thioesterases. Structural comparison of FlK complexed with various substrate analogues suggests that the interaction between the fluorine of the substrate and the side chain of Arg(120) located opposite to the catalytic triad is essential for correct coordination of the substrate at the active site and therefore accounts for the substrate specificity.


  • Organizational Affiliation

    Department of Biochemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, United Kingdom.


Macromolecules
Find similar proteins by:  (by identity cutoff)  |  3D Structure
Entity ID: 1
MoleculeChains Sequence LengthOrganismDetailsImage
Fluoroacetyl-CoA thioesterase FlK
A, B, C, D, E
A, B, C, D, E, F, G, H
139Streptantibioticus cattleyicolorMutation(s): 0 
Gene Names: flKfluoroacetyl-CoA thioesterase FlK
EC: 3
UniProt
Find proteins for Q1EMV2 (Streptantibioticus cattleyicolor)
Explore Q1EMV2 
Go to UniProtKB:  Q1EMV2
Entity Groups  
Sequence Clusters30% Identity50% Identity70% Identity90% Identity95% Identity100% Identity
UniProt GroupQ1EMV2
Sequence Annotations
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  • Reference Sequence
Small Molecules
Ligands 1 Unique
IDChains Name / Formula / InChI Key2D Diagram3D Interactions
ENV
Query on ENV

Download Ideal Coordinates CCD File 
I [auth B],
J [auth E]
(2R)-N-{3-[(5-fluoro-4-oxopentyl)amino]-3-oxopropyl}-2,4-dihydroxy-3,3-dimethylbutanamide
C14 H25 F N2 O5
FTHCNOQGZZPFFG-LBPRGKRZSA-N
Experimental Data & Validation

Experimental Data

  • Method: X-RAY DIFFRACTION
  • Resolution: 2.10 Å
  • R-Value Free: 0.282 
  • R-Value Work: 0.204 
  • R-Value Observed: 0.208 
  • Space Group: P 1 21 1
Unit Cell:
Length ( Å )Angle ( ˚ )
a = 79.5α = 90
b = 70.89β = 103.06
c = 102.88γ = 90
Software Package:
Software NamePurpose
MAR345dtbdata collection
AMoREphasing
REFMACrefinement
MOSFLMdata reduction
SCALAdata scaling

Structure Validation

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Ligand Structure Quality Assessment 


Entry History 

Deposition Data

Revision History  (Full details and data files)

  • Version 1.0: 2010-04-28
    Type: Initial release
  • Version 1.1: 2011-07-13
    Changes: Version format compliance
  • Version 1.2: 2018-01-24
    Changes: Advisory, Structure summary
  • Version 1.3: 2023-09-06
    Changes: Advisory, Data collection, Database references, Derived calculations, Refinement description