Hydrolytic catalysis and structural stabilization in a designed metalloprotein

Metal ions are an important part of many natural proteins, providing structural, catalytic and electron transfer functions. Reproducing these functions in a designed protein is the ultimate challenge to our understanding of them. Here, we present an artificial metallohydrolase, which has been shown...

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Main Authors:	Zastrow, Melissa L., Peacock, Anna F. A., Stuckey, Jeanne A., Pecoraro, Vincent L.
Format:	Online
Language:	English
Published:	2011
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3270697/

id	pubmed-3270697
recordtype	oai_dc
spelling	pubmed-32706972012-08-01 Hydrolytic catalysis and structural stabilization in a designed metalloprotein Zastrow, Melissa L. Peacock, Anna F. A. Stuckey, Jeanne A. Pecoraro, Vincent L. Article Metal ions are an important part of many natural proteins, providing structural, catalytic and electron transfer functions. Reproducing these functions in a designed protein is the ultimate challenge to our understanding of them. Here, we present an artificial metallohydrolase, which has been shown by X-ray crystallography to contain two different metal ions – a Zn(II) ion which is important for catalytic activity and a Hg(II) ion which provides structural stability. This metallohydrolase displays catalytic activity that compares well with several characteristic reactions of natural enzymes. It catalyses p-nitrophenyl acetate hydrolysis (pNPA) to within ~100-fold of the efficiency of human carbonic anhydrase (CA)II and is at least 550-fold better than comparable synthetic complexes. Similarly, CO2 hydration occurs with an efficiency within ~500-fold of CAII. While histidine residues in the absence of Zn(II) exhibit pNPA hydrolysis, miniscule apopeptide activity is observed for CO2 hydration. The kinetic and structural analysis of this first de novo designed hydrolytic metalloenzyme uncovers necessary design features for future metalloenzymes containing one or more metals. 2011-11-27 /pmc/articles/PMC3270697/ /pubmed/22270627 http://dx.doi.org/10.1038/nchem.1201 Text en Users may view, print, copy, download and text and data- mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use: http://www.nature.com/authors/editorial_policies/license.html#terms
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Zastrow, Melissa L. Peacock, Anna F. A. Stuckey, Jeanne A. Pecoraro, Vincent L.
spellingShingle	Zastrow, Melissa L. Peacock, Anna F. A. Stuckey, Jeanne A. Pecoraro, Vincent L. Hydrolytic catalysis and structural stabilization in a designed metalloprotein
author_facet	Zastrow, Melissa L. Peacock, Anna F. A. Stuckey, Jeanne A. Pecoraro, Vincent L.
author_sort	Zastrow, Melissa L.
title	Hydrolytic catalysis and structural stabilization in a designed metalloprotein
title_short	Hydrolytic catalysis and structural stabilization in a designed metalloprotein
title_full	Hydrolytic catalysis and structural stabilization in a designed metalloprotein
title_fullStr	Hydrolytic catalysis and structural stabilization in a designed metalloprotein
title_full_unstemmed	Hydrolytic catalysis and structural stabilization in a designed metalloprotein
title_sort	hydrolytic catalysis and structural stabilization in a designed metalloprotein
description	Metal ions are an important part of many natural proteins, providing structural, catalytic and electron transfer functions. Reproducing these functions in a designed protein is the ultimate challenge to our understanding of them. Here, we present an artificial metallohydrolase, which has been shown by X-ray crystallography to contain two different metal ions – a Zn(II) ion which is important for catalytic activity and a Hg(II) ion which provides structural stability. This metallohydrolase displays catalytic activity that compares well with several characteristic reactions of natural enzymes. It catalyses p-nitrophenyl acetate hydrolysis (pNPA) to within ~100-fold of the efficiency of human carbonic anhydrase (CA)II and is at least 550-fold better than comparable synthetic complexes. Similarly, CO2 hydration occurs with an efficiency within ~500-fold of CAII. While histidine residues in the absence of Zn(II) exhibit pNPA hydrolysis, miniscule apopeptide activity is observed for CO2 hydration. The kinetic and structural analysis of this first de novo designed hydrolytic metalloenzyme uncovers necessary design features for future metalloenzymes containing one or more metals.
publishDate	2011
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3270697/
_version_	1611503337688006656

Hydrolytic catalysis and structural stabilization in a designed metalloprotein

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