Issue 32, 2018

Engineering a bifunctional copper site in the cupredoxin fold by loop-directed mutagenesis

Abstract

Copper sites in proteins are designed to perform either electron transfer or redox catalysis. Type 1 and CuA sites are electron transfer hubs bound to a rigid protein fold that prevents binding of exogenous ligands and side reactions. Here we report the engineering of two Type 1 sites by loop-directed mutagenesis within a CuA scaffold with unique electronic structures and functional features. A copper–thioether axial bond shorter than the copper–thiolate bond is responsible for the electronic structure features, in contrast to all other natural or chimeric sites where the copper thiolate bond is short. These sites display highly unusual features, such as: (1) a high reduction potential despite a strong interaction with the axial ligand, which we attribute to changes in the hydrogen bond network and (2) the ability to bind exogenous ligands such as imidazole and azide. This strategy widens the possibility of using natural protein scaffolds with functional features not present in nature.

Graphical abstract: Engineering a bifunctional copper site in the cupredoxin fold by loop-directed mutagenesis

Supplementary files

Article information

Article type
Edge Article
Submitted
29 Marts 2018
Accepted
27 Jūn. 2018
First published
28 Jūn. 2018
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., 2018,9, 6692-6702

Engineering a bifunctional copper site in the cupredoxin fold by loop-directed mutagenesis

A. Espinoza-Cara, U. Zitare, D. Alvarez-Paggi, S. Klinke, L. H. Otero, D. H. Murgida and A. J. Vila, Chem. Sci., 2018, 9, 6692 DOI: 10.1039/C8SC01444B

This article is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported Licence. You can use material from this article in other publications, without requesting further permission from the RSC, provided that the correct acknowledgement is given and it is not used for commercial purposes.

To request permission to reproduce material from this article in a commercial publication, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party commercial publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements