A conserved threonine prevents self-intoxication of enoyl-thioester reductases

Nature Chemical Biology
Raoul G RosenthalTobias J Erb

Abstract

Enzymes are highly specific biocatalysts, yet they can promote unwanted side reactions. Here we investigated the factors that direct catalysis in the enoyl-thioester reductase Etr1p. We show that a single conserved threonine is essential to suppress the formation of a side product that would otherwise act as a high-affinity inhibitor of the enzyme. Substitution of this threonine with isosteric valine increases side-product formation by more than six orders of magnitude, while decreasing turnover frequency by only one order of magnitude. Our results show that the promotion of wanted reactions and the suppression of unwanted side reactions operate independently at the active site of Etr1p, and that the active suppression of side reactions is highly conserved in the family of medium-chain dehydrogenases/reductases (MDRs). Our discovery emphasizes the fact that the active destabilization of competing transition states is an important factor during catalysis that has implications for the understanding and the de novo design of enzymes.

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Citations

Oct 14, 2017·Science·Stephan C HammerFrances H Arnold
Sep 16, 2018·The Journal of Biological Chemistry·Bastian VögeliTobias J Erb
Jan 29, 2021·Current Opinion in Biotechnology·Iria BernhardsgrütterTobias J Erb
Jun 23, 2021·Chemical Reviews·Pedro PaivaPedro A Fernandes
Jun 13, 2019·Journal of the American Chemical Society·Iria BernhardsgrütterTobias J Erb
Mar 3, 2018·ACS Synthetic Biology·Nicolas Krink-KoutsoubelisJay D Keasling

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