4F21

Crystal structure of carboxylesterase/phospholipase family protein from Francisella tularensis


Experimental Data Snapshot

  • Method: X-RAY DIFFRACTION
  • Resolution: 2.50 Å
  • R-Value Free: 0.259 
  • R-Value Work: 0.199 
  • R-Value Observed: 0.204 

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Ligand Structure Quality Assessment 


This is version 1.4 of the entry. See complete history


Literature

Large scale structural rearrangement of a serine hydrolase from Francisella tularensis facilitates catalysis.

Filippova, E.V.Weston, L.A.Kuhn, M.L.Geissler, B.Gehring, A.M.Armoush, N.Adkins, C.T.Minasov, G.Dubrovska, I.Shuvalova, L.Winsor, J.R.Lavis, L.D.Satchell, K.J.Becker, D.P.Anderson, W.F.Johnson, R.J.

(2013) J Biol Chem 288: 10522-10535

  • DOI: https://doi.org/10.1074/jbc.M112.446625
  • Primary Citation of Related Structures:  
    4F21

  • PubMed Abstract: 

    Tularemia is a deadly, febrile disease caused by infection by the gram-negative bacterium, Francisella tularensis. Members of the ubiquitous serine hydrolase protein family are among current targets to treat diverse bacterial infections. Herein we present a structural and functional study of a novel bacterial carboxylesterase (FTT258) from F. tularensis, a homologue of human acyl protein thioesterase (hAPT1). The structure of FTT258 has been determined in multiple forms, and unexpectedly large conformational changes of a peripheral flexible loop occur in the presence of a mechanistic cyclobutanone ligand. The concomitant changes in this hydrophobic loop and the newly exposed hydrophobic substrate binding pocket suggest that the observed structural changes are essential to the biological function and catalytic activity of FTT258. Using diverse substrate libraries, site-directed mutagenesis, and liposome binding assays, we determined the importance of these structural changes to the catalytic activity and membrane binding activity of FTT258. Residues within the newly exposed hydrophobic binding pocket and within the peripheral flexible loop proved essential to the hydrolytic activity of FTT258, indicating that structural rearrangement is required for catalytic activity. Both FTT258 and hAPT1 also showed significant association with liposomes designed to mimic bacterial or human membranes, respectively, even though similar structural rearrangements for hAPT1 have not been reported. The necessity for acyl protein thioesterases to have maximal catalytic activity near the membrane surface suggests that these conformational changes in the protein may dually regulate catalytic activity and membrane association in bacterial and human homologues.


  • Organizational Affiliation

    Center for Structural Genomics of Infectious Diseases and the Department of Molecular Pharmacology and Biological Chemistry, Northwestern University Feinberg School of Medicine, Chicago, Illinois 60611, USA.


Macromolecules
Find similar proteins by:  (by identity cutoff)  |  3D Structure
Entity ID: 1
MoleculeChains Sequence LengthOrganismDetailsImage
Carboxylesterase/phospholipase family protein
A, B, C, D, E
A, B, C, D, E, F, G, H
246Francisella tularensis subsp. tularensis SCHU S4Mutation(s): 0 
Gene Names: FTT_0258
EC: 3.1.1
UniProt
Find proteins for Q5NI32 (Francisella tularensis subsp. tularensis (strain SCHU S4 / Schu 4))
Explore Q5NI32 
Go to UniProtKB:  Q5NI32
Entity Groups  
Sequence Clusters30% Identity50% Identity70% Identity90% Identity95% Identity100% Identity
UniProt GroupQ5NI32
Sequence Annotations
Expand
  • Reference Sequence
Small Molecules
Ligands 1 Unique
IDChains Name / Formula / InChI Key2D Diagram3D Interactions
0S1
Query on 0S1

Download Ideal Coordinates CCD File 
I [auth E]N-((1R,2S)-2-allyl-4-oxocyclobutyl)-4-methylbenzenesulfonamide, bound form
C14 H19 N O3 S
KFIWUGSQQLTLIY-FPMFFAJLSA-N
Experimental Data & Validation

Experimental Data

  • Method: X-RAY DIFFRACTION
  • Resolution: 2.50 Å
  • R-Value Free: 0.259 
  • R-Value Work: 0.199 
  • R-Value Observed: 0.204 
  • Space Group: P 1
  • Diffraction Data: https://doi.org/10.18430/M34F21
Unit Cell:
Length ( Å )Angle ( ˚ )
a = 50.997α = 94.89
b = 64.416β = 90.12
c = 139.039γ = 89.96
Software Package:
Software NamePurpose
Blu-Icedata collection
MOLREPphasing
PHENIXrefinement
HKL-2000data reduction
HKL-2000data scaling

Structure Validation

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Ligand Structure Quality Assessment 


Entry History 

Revision History  (Full details and data files)

  • Version 1.0: 2012-08-29
    Type: Initial release
  • Version 1.1: 2013-10-09
    Changes: Database references
  • Version 1.2: 2017-11-15
    Changes: Advisory, Refinement description
  • Version 1.3: 2018-01-24
    Changes: Structure summary
  • Version 1.4: 2023-09-13
    Changes: Advisory, Data collection, Database references, Derived calculations, Refinement description