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Nat Chem Biol. 2017 Jul;13(7):745-749. doi: 10.1038/nchembio.2375. Epub 2017 May 15.

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

Author information

1
Biochemistry and Synthetic Biology of Microbial Metabolism Group, Max Planck Institute for Terrestrial Microbiology, Marburg, Germany.
2
Microbial Protein Structure Group, Max Planck Institute for Terrestrial Microbiology, Marburg, Germany.

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.

PMID:
28504678
PMCID:
PMC5892710
DOI:
10.1038/nchembio.2375
[Indexed for MEDLINE]
Free PMC Article

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