The evolution of multiple active site configurations in a designed enzyme.

Autor:	Hong NS; Research School of Chemistry, Australian National University, Canberra, ACT, 2601, Australia., Petrović D; Department of Chemistry, BMC, Uppsala University, Box 576, 751 23, Uppsala, Sweden., Lee R; Research School of Chemistry, Australian National University, Canberra, ACT, 2601, Australia., Gryn'ova G; Research School of Chemistry, Australian National University, Canberra, ACT, 2601, Australia.; Institut des Sciences et Ingénierie Chimiques, École Polytechnique Fédérale de Lausanne, 1015, Lausanne, Switzerland., Purg M; Department of Chemistry, BMC, Uppsala University, Box 576, 751 23, Uppsala, Sweden., Saunders J; Research School of Chemistry, Australian National University, Canberra, ACT, 2601, Australia., Bauer P; Department of Chemistry, BMC, Uppsala University, Box 576, 751 23, Uppsala, Sweden., Carr PD; Research School of Chemistry, Australian National University, Canberra, ACT, 2601, Australia., Lin CY; Research School of Chemistry, Australian National University, Canberra, ACT, 2601, Australia., Mabbitt PD; Research School of Chemistry, Australian National University, Canberra, ACT, 2601, Australia., Zhang W; Research School of Chemistry, Australian National University, Canberra, ACT, 2601, Australia., Altamore T; Research School of Chemistry, Australian National University, Canberra, ACT, 2601, Australia., Easton C; Research School of Chemistry, Australian National University, Canberra, ACT, 2601, Australia., Coote ML; Research School of Chemistry, Australian National University, Canberra, ACT, 2601, Australia., Kamerlin SCL; Department of Chemistry, BMC, Uppsala University, Box 576, 751 23, Uppsala, Sweden. lynn.kamerlin@kemi.uu.se., Jackson CJ; Research School of Chemistry, Australian National University, Canberra, ACT, 2601, Australia. colin.jackson@anu.edu.au.
Jazyk:	angličtina
Zdroj:	Nature communications [Nat Commun] 2018 Sep 25; Vol. 9 (1), pp. 3900. Date of Electronic Publication: 2018 Sep 25.
DOI:	10.1038/s41467-018-06305-y
Abstrakt:	Developments in computational chemistry, bioinformatics, and laboratory evolution have facilitated the de novo design and catalytic optimization of enzymes. Besides creating useful catalysts, the generation and iterative improvement of designed enzymes can provide valuable insight into the interplay between the many phenomena that have been suggested to contribute to catalysis. In this work, we follow changes in conformational sampling, electrostatic preorganization, and quantum tunneling along the evolutionary trajectory of a designed Kemp eliminase. We observe that in the Kemp Eliminase KE07, instability of the designed active site leads to the emergence of two additional active site configurations. Evolutionary conformational selection then gradually stabilizes the most efficient configuration, leading to an improved enzyme. This work exemplifies the link between conformational plasticity and evolvability and demonstrates that residues remote from the active sites of enzymes play crucial roles in controlling and shaping the active site for efficient catalysis.
Databáze:	MEDLINE
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