Issue 10, 2017

Design of an enantioselective artificial metallo-hydratase enzyme containing an unnatural metal-binding amino acid

Abstract

The design of artificial metalloenzymes is a challenging, yet ultimately highly rewarding objective because of the potential for accessing new-to-nature reactions. One of the main challenges is identifying catalytically active substrate–metal cofactor–host geometries. The advent of expanded genetic code methods for the in vivo incorporation of non-canonical metal-binding amino acids into proteins allow to address an important aspect of this challenge: the creation of a stable, well-defined metal-binding site. Here, we report a designed artificial metallohydratase, based on the transcriptional repressor lactococcal multidrug resistance regulator (LmrR), in which the non-canonical amino acid (2,2′-bipyridin-5yl)alanine is used to bind the catalytic Cu(II) ion. Starting from a set of empirical pre-conditions, a combination of cluster model calculations (QM), protein–ligand docking and molecular dynamics simulations was used to propose metallohydratase variants, that were experimentally verified. The agreement observed between the computationally predicted and experimentally observed catalysis results demonstrates the power of the artificial metalloenzyme design approach presented here.

Graphical abstract: Design of an enantioselective artificial metallo-hydratase enzyme containing an unnatural metal-binding amino acid

Supplementary files

Article information

Article type
Edge Article
Submitted
09 Aug 2017
Accepted
01 Sep 2017
First published
04 Sep 2017
This article is Open Access

All publication charges for this article have been paid for by the Royal Society of Chemistry
Creative Commons BY-NC license

Chem. Sci., 2017,8, 7228-7235

Design of an enantioselective artificial metallo-hydratase enzyme containing an unnatural metal-binding amino acid

I. Drienovská, L. Alonso-Cotchico, P. Vidossich, A. Lledós, J. Maréchal and G. Roelfes, Chem. Sci., 2017, 8, 7228 DOI: 10.1039/C7SC03477F

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