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In Vitro Reconstitution of Formylglycine-Generating Enzymes Requires Copper(I)

Knop, Matthias and Engi, Pascal and Lemnaru, Roxana and Seebeck, Florian P.. (2015) In Vitro Reconstitution of Formylglycine-Generating Enzymes Requires Copper(I). ChemBioChem, 16 (15). pp. 2147-2150.

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Official URL: http://edoc.unibas.ch/41782/

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Abstract

Formylglycine-generating enzymes (FGEs) catalyze O2 -dependent conversion of specific cysteine residues of arylsulfatases and alkaline phosphatases into formylglycine. The ability also to introduce unique aldehyde functions into recombinant proteins makes FGEs a powerful tool for protein engineering. One limitation of this technology is poor in vitro activity of reconstituted FGEs. Although FGEs have been characterized as cofactor-free enzymes we report that the addition of one equivalent of Cu(I) increases catalytic efficiency more than 20-fold and enables the identification of stereoselective C-H bond cleavage at the substrate as the rate-limiting step. These findings remove previous limitations of FGE-based protein engineering and also pose new questions about the catalytic mechanism of this O2 -utilizing enzyme.
Faculties and Departments:05 Faculty of Science > Departement Chemie > Chemie > Molecular Bionics (Seebeck)
UniBasel Contributors:Seebeck, Florian Peter
Item Type:Article, refereed
Article Subtype:Research Article
Publisher:Wiley
ISSN:1439-4227
e-ISSN:1439-7633
Note:Publication type according to Uni Basel Research Database: Journal article
Identification Number:
Last Modified:12 Apr 2017 09:28
Deposited On:25 Apr 2016 13:16

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