7K4U
Crystal structure of Kemp Eliminase HG3 K50Q in complex with the transition state analog 6-nitrobenzotriazole
Summary for 7K4U
| Entry DOI | 10.2210/pdb7k4u/pdb |
| Descriptor | Endo-1,4-beta-xylanase, 6-NITROBENZOTRIAZOLE (3 entities in total) |
| Functional Keywords | kemp elimination, directed evolution, transition state analog, hydrolase |
| Biological source | Thermoascus aurantiacus |
| Total number of polymer chains | 1 |
| Total formula weight | 33235.29 |
| Authors | Padua, R.A.P.,Otten, R.,Bunzel, A.,Nguyen, V.,Pitsawong, W.,Patterson, M.,Sui, S.,Perry, S.L.,Cohen, A.E.,Hilvert, D.,Kern, D. (deposition date: 2020-09-16, release date: 2020-12-02, Last modification date: 2023-10-18) |
| Primary citation | Otten, R.,Padua, R.A.P.,Bunzel, H.A.,Nguyen, V.,Pitsawong, W.,Patterson, M.,Sui, S.,Perry, S.L.,Cohen, A.E.,Hilvert, D.,Kern, D. How directed evolution reshapes the energy landscape in an enzyme to boost catalysis. Science, 370:1442-1446, 2020 Cited by PubMed Abstract: The advent of biocatalysts designed computationally and optimized by laboratory evolution provides an opportunity to explore molecular strategies for augmenting catalytic function. Applying a suite of nuclear magnetic resonance, crystallography, and stopped-flow techniques to an enzyme designed for an elementary proton transfer reaction, we show how directed evolution gradually altered the conformational ensemble of the protein scaffold to populate a narrow, highly active conformational ensemble and accelerate this transformation by nearly nine orders of magnitude. Mutations acquired during optimization enabled global conformational changes, including high-energy backbone rearrangements, that cooperatively organized the catalytic base and oxyanion stabilizer, thus perfecting transition-state stabilization. The development of protein catalysts for many chemical transformations could be facilitated by explicitly sampling conformational substates during design and specifically stabilizing productive substates over all unproductive conformations. PubMed: 33214289DOI: 10.1126/science.abd3623 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.3 Å) |
Structure validation
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