4OYL

Humicola insolens cutinase in complex with mono-ethylphosphate


Experimental Data Snapshot

  • Method: X-RAY DIFFRACTION
  • Resolution: 2.05 Å
  • R-Value Free: 0.184 
  • R-Value Work: 0.143 
  • R-Value Observed: 0.145 

wwPDB Validation   3D Report Full Report


This is version 1.5 of the entry. See complete history


Literature

Thermodynamic and structural investigation of the specific SDS binding of Humicola insolens cutinase.

Kold, D.Dauter, Z.Laustsen, A.K.Brzozowski, A.M.Turkenburg, J.P.Nielsen, A.D.Kolds, H.Petersen, E.Schitt, B.De Maria, L.Wilson, K.S.Svendsen, A.Wimmer, R.

(2014) Protein Sci 23: 1023-1035

  • DOI: https://doi.org/10.1002/pro.2489
  • Primary Citation of Related Structures:  
    4OYL, 4OYY

  • PubMed Abstract: 

    The interaction of lipolytic enzymes with anionic surfactants is of great interest with respect to industrially produced detergents. Here, we report the interaction of cutinase from the thermophilic fungus Humicola insolens with the anionic surfactant SDS, and show the enzyme specifically binds a single SDS molecule under nondenaturing concentrations. Protein interaction with SDS was investigated by NMR, ITC and molecular dynamics simulations. The NMR resonances of the protein were assigned, with large stretches of the protein molecule not showing any detectable resonances. SDS is shown to specifically interact with the loops surrounding the catalytic triad with medium affinity (Ka ≈ 10(5) M(-1) ). The mode of binding is closely similar to that seen previously for binding of amphiphilic molecules and substrate analogues to cutinases, and hence SDS acts as a substrate mimic. In addition, the structure of the enzyme has been solved by X-ray crystallography in its apo form and after cocrystallization with diethyl p-nitrophenyl phosphate (DNPP) leading to a complex with monoethylphosphate (MEP) esterified to the catalytically active serine. The enzyme has the same fold as reported for other cutinases but, unexpectedly, esterification of the active site serine is accompanied by the ethylation of the active site histidine which flips out from its usual position in the triad.


  • Organizational Affiliation

    Department of Biotechnology, Chemistry and Environmental Engineering, Aalborg University, Sohngaardsholmsvej 49, DK-9000, Aalborg, Denmark.


Macromolecules
Find similar proteins by:  (by identity cutoff)  |  3D Structure
Entity ID: 1
MoleculeChains Sequence LengthOrganismDetailsImage
Cutinase
A, B, C
194Humicola insolensMutation(s): 0 
UniProt
Find proteins for A0A075B5G4 (Humicola insolens)
Explore A0A075B5G4 
Go to UniProtKB:  A0A075B5G4
Entity Groups  
Sequence Clusters30% Identity50% Identity70% Identity90% Identity95% Identity100% Identity
UniProt GroupA0A075B5G4
Sequence Annotations
Expand
  • Reference Sequence
Small Molecules
Modified Residues  2 Unique
IDChains TypeFormula2D DiagramParent
MIR
Query on MIR
A, B, C
L-PEPTIDE LINKINGC5 H12 N O6 PSER
OYL
Query on OYL
A, B, C
L-PEPTIDE LINKINGC8 H13 N3 O2HIS
Experimental Data & Validation

Experimental Data

  • Method: X-RAY DIFFRACTION
  • Resolution: 2.05 Å
  • R-Value Free: 0.184 
  • R-Value Work: 0.143 
  • R-Value Observed: 0.145 
  • Space Group: P 1 21 1
Unit Cell:
Length ( Å )Angle ( ˚ )
a = 71.63α = 90
b = 66.4β = 119.3
c = 71.98γ = 90
Software Package:
Software NamePurpose
DENZOdata reduction
SCALEPACKdata scaling
AMoREphasing
REFMACrefinement

Structure Validation

View Full Validation Report



Entry History 

Deposition Data

Revision History  (Full details and data files)

  • Version 1.0: 2014-06-25
    Type: Initial release
  • Version 1.1: 2014-10-01
    Changes: Database references
  • Version 1.2: 2015-01-07
    Changes: Database references
  • Version 1.3: 2015-09-23
    Changes: Data collection
  • Version 1.4: 2017-11-22
    Changes: Data collection, Database references, Derived calculations, Refinement description
  • Version 1.5: 2023-12-27
    Changes: Data collection, Database references, Derived calculations