3T1G
Engineering of organophosphate hydrolase by computational design and directed evolution
Summary for 3T1G
Entry DOI | 10.2210/pdb3t1g/pdb |
Related | 1A4L |
Descriptor | organophosphate hydrolase, ZINC ION, CALCIUM ION, ... (4 entities in total) |
Functional Keywords | computational design, directed evolution, tim beta/alpha-barrel, metallo-dependent hydrolase, organophosphate binding, hydrolysis, artificial enzyme, hydrolase |
Biological source | Mus musculus (mouse) |
Cellular location | Cell membrane; Peripheral membrane protein; Extracellular side (By similarity): P03958 |
Total number of polymer chains | 1 |
Total formula weight | 40482.39 |
Authors | Takeuchi, R.,Stoddard, B.L. (deposition date: 2011-07-21, release date: 2012-02-08, Last modification date: 2023-09-13) |
Primary citation | Khare, S.D.,Kipnis, Y.,Greisen, P.J.,Takeuchi, R.,Ashani, Y.,Goldsmith, M.,Song, Y.,Gallaher, J.L.,Silman, I.,Leader, H.,Sussman, J.L.,Stoddard, B.L.,Tawfik, D.S.,Baker, D. Computational redesign of a mononuclear zinc metalloenzyme for organophosphate hydrolysis. Nat.Chem.Biol., 8:294-300, 2012 Cited by PubMed Abstract: 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. PubMed: 22306579DOI: 10.1038/nchembio.777 PDB entries with the same primary citation |
Experimental method | X-RAY DIFFRACTION (2.35 Å) |
Structure validation
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