A biosynthetic approach to understanding the assembly of CuA centers
type: CreativeWork
description: Electron transfer plays a critical role in countless biological processes. Metalloproteins containing copper or iron carry out electron transfer throughout all domains of life. CuA is a unique dinuclear, mixed-valence and valence delocalized copper center, which catalyzes remarkably efficient electron transfer under the low driving forces experienced at the termini of electron transport chains. Historically, CuA centers have been difficult to study, due to their occurrence in large enzyme complexes that are embedded in membranes and/or the presence of other metallocofactors in the same enzyme. For these reasons, soluble proteins containing only the CuA cofactor were created, either by truncating helices that anchored the CuA domain to a membrane, or by engineering the CuA center into another protein. The strategy of engineering CuA centers into another protein falls under the umbrella of the biosynthetic approach to metalloenzyme studies. In the biosynthetic approach to metalloenzyme studies, the protein is considered to be a large ligand to the metal center, which is customizable to the needs of that center. The biosynthetic approach offers many distinctive benefits in the study of native metal centers over the complementary approach of studying these centers in their native enzymes. These benefits include simpler protein systems, which are easy to express and purify in high yield and are free of other metal cofactors, and the ability to substitute non-native metals into the site. A biosynthetic model of CuA centers was previously designed into a mononuclear copper enzyme called azurin (Az). In this model, the CuA center was engineered into the same location of the protein as the original copper center, by replacement of a single loop containing a majority of the copper ligands. The resulting CuAAz construct binds copper in a copper site that is very similar to native CuA centers, as evidenced by many spectroscopic and structural studies. The ease of purification, …
abstract: Electron transfer plays a critical role in countless biological processes. Metalloproteins containing copper or iron carry out electron transfer throughout all domains of life. CuA is a unique dinuclear, mixed-valence and valence delocalized copper center, which catalyzes remarkably efficient electron transfer under the low driving forces experienced at the termini of electron transport chains. Historically, CuA centers have been difficult to study, due to their occurrence in large enzyme complexes that are embedded in membranes and/or the presence of other metallocofactors in the same enzyme. For these reasons, soluble proteins containing only the CuA cofactor were created, either by truncating helices that anchored the CuA domain to a membrane, or by engineering the CuA center into another protein. The strategy of engineering CuA centers into another protein falls under the umbrella of the biosynthetic approach to metalloenzyme studies. In the biosynthetic approach to metalloenzyme studies, the protein is considered to be a large ligand to the metal center, which is customizable to the needs of that center. The biosynthetic approach offers many distinctive benefits in the study of native metal centers over the complementary approach of studying these centers in their native enzymes. These benefits include simpler protein systems, which are easy to express and purify in high yield and are free of other metal cofactors, and the ability to substitute non-native metals into the site. A biosynthetic model of CuA centers was previously designed into a mononuclear copper enzyme called azurin (Az). In this model, the CuA center was engineered into the same location of the protein as the original copper center, by replacement of a single loop containing a majority of the copper ligands. The resulting CuAAz construct binds copper in a copper site that is very similar to native CuA centers, as evidenced by many spectroscopic and structural studies. The ease of purification, …
author: Wilson, Tiffany
language: en
keywords: biological electron transfer, CuA, CuA assembly, Sco, copper chaperones, metalloprotein design, azurin
date published: 2012-09-18T21:24:39Z
date modified: 2026-07-22T22:25:31Z
provider: University of Illinois - Urbana-Champaign
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: University of Illinois at Urbana-Champaign

PropertyValue
property ID: Handle
value: http://hdl.handle.net/2142/34537