Computational redesign of a mononuclear zinc metalloenzyme for organophosphate hydrolysis

Nat Chem Biol. 2012 Feb 5;8(3):294-300. doi: 10.1038/nchembio.777.


The ability to redesign enzymes to catalyze noncognate chemical transformations would have wide-ranging applications. We developed a computational method for repurposing the reactivity of metalloenzyme active site functional groups to catalyze new reactions. Using this method, we engineered a zinc-containing mouse adenosine deaminase to catalyze the hydrolysis of a model organophosphate with a catalytic efficiency (k(cat)/K(m)) of ~10(4) M(-1) s(-1) after directed evolution. In the high-resolution crystal structure of the enzyme, all but one of the designed residues adopt the designed conformation. The designed enzyme efficiently catalyzes the hydrolysis of the R(P) isomer of a coumarinyl analog of the nerve agent cyclosarin, and it shows marked substrate selectivity for coumarinyl leaving groups. Computational redesign of native enzyme active sites complements directed evolution methods and offers a general approach for exploring their untapped catalytic potential for new reactivities.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Adenosine Deaminase / chemistry
  • Adenosine Deaminase / metabolism*
  • Animals
  • Biocatalysis
  • Catalytic Domain
  • Computational Biology
  • Computer Simulation*
  • Computer-Aided Design*
  • Hydrolysis
  • Metalloproteins / chemistry
  • Metalloproteins / metabolism*
  • Mice
  • Models, Molecular
  • Molecular Conformation
  • Organophosphorus Compounds / chemistry
  • Organophosphorus Compounds / metabolism*
  • Zinc / chemistry*
  • Zinc / metabolism


  • Metalloproteins
  • Organophosphorus Compounds
  • Adenosine Deaminase
  • Zinc

Associated data

  • PDB/3T1G