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

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-direct...

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Autores principales:	Espinoza-Cara, A., Zitare, U., Alvarez-Paggi, D., Klinke, S., Otero, L.H., Murgida, D.H., Vila, A.J.
Formato:	JOUR
Materias:	Binding sites Electron transitions Electronic structure Hydrogen bonds Ligands Mutagenesis Proteins Scaffolds (biology) Directed mutagenesis Electron transfer Exogenous ligands Functional features Hydrogen bond networks Natural proteins Reduction potential Strong interaction Copper
Acceso en línea:	http://hdl.handle.net/20.500.12110/paper_20416520_v9_n32_p6692_EspinozaCara
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	todo:paper_20416520_v9_n32_p6692_EspinozaCara
record_format	dspace
spelling	todo:paper_20416520_v9_n32_p6692_EspinozaCara2023-10-03T16:37:58Z Engineering a bifunctional copper site in the cupredoxin fold by loop-directed mutagenesis Espinoza-Cara, A. Zitare, U. Alvarez-Paggi, D. Klinke, S. Otero, L.H. Murgida, D.H. Vila, A.J. Binding sites Electron transitions Electronic structure Hydrogen bonds Ligands Mutagenesis Proteins Scaffolds (biology) Directed mutagenesis Electron transfer Exogenous ligands Functional features Hydrogen bond networks Natural proteins Reduction potential Strong interaction Copper 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. © The Royal Society of Chemistry. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_20416520_v9_n32_p6692_EspinozaCara
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	Binding sites Electron transitions Electronic structure Hydrogen bonds Ligands Mutagenesis Proteins Scaffolds (biology) Directed mutagenesis Electron transfer Exogenous ligands Functional features Hydrogen bond networks Natural proteins Reduction potential Strong interaction Copper
spellingShingle	Binding sites Electron transitions Electronic structure Hydrogen bonds Ligands Mutagenesis Proteins Scaffolds (biology) Directed mutagenesis Electron transfer Exogenous ligands Functional features Hydrogen bond networks Natural proteins Reduction potential Strong interaction Copper Espinoza-Cara, A. Zitare, U. Alvarez-Paggi, D. Klinke, S. Otero, L.H. Murgida, D.H. Vila, A.J. Engineering a bifunctional copper site in the cupredoxin fold by loop-directed mutagenesis
topic_facet	Binding sites Electron transitions Electronic structure Hydrogen bonds Ligands Mutagenesis Proteins Scaffolds (biology) Directed mutagenesis Electron transfer Exogenous ligands Functional features Hydrogen bond networks Natural proteins Reduction potential Strong interaction Copper
description	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. © The Royal Society of Chemistry.
format	JOUR
author	Espinoza-Cara, A. Zitare, U. Alvarez-Paggi, D. Klinke, S. Otero, L.H. Murgida, D.H. Vila, A.J.
author_facet	Espinoza-Cara, A. Zitare, U. Alvarez-Paggi, D. Klinke, S. Otero, L.H. Murgida, D.H. Vila, A.J.
author_sort	Espinoza-Cara, A.
title	Engineering a bifunctional copper site in the cupredoxin fold by loop-directed mutagenesis
title_short	Engineering a bifunctional copper site in the cupredoxin fold by loop-directed mutagenesis
title_full	Engineering a bifunctional copper site in the cupredoxin fold by loop-directed mutagenesis
title_fullStr	Engineering a bifunctional copper site in the cupredoxin fold by loop-directed mutagenesis
title_full_unstemmed	Engineering a bifunctional copper site in the cupredoxin fold by loop-directed mutagenesis
title_sort	engineering a bifunctional copper site in the cupredoxin fold by loop-directed mutagenesis
url	http://hdl.handle.net/20.500.12110/paper_20416520_v9_n32_p6692_EspinozaCara
work_keys_str_mv	AT espinozacaraa engineeringabifunctionalcoppersiteinthecupredoxinfoldbyloopdirectedmutagenesis AT zitareu engineeringabifunctionalcoppersiteinthecupredoxinfoldbyloopdirectedmutagenesis AT alvarezpaggid engineeringabifunctionalcoppersiteinthecupredoxinfoldbyloopdirectedmutagenesis AT klinkes engineeringabifunctionalcoppersiteinthecupredoxinfoldbyloopdirectedmutagenesis AT oterolh engineeringabifunctionalcoppersiteinthecupredoxinfoldbyloopdirectedmutagenesis AT murgidadh engineeringabifunctionalcoppersiteinthecupredoxinfoldbyloopdirectedmutagenesis AT vilaaj engineeringabifunctionalcoppersiteinthecupredoxinfoldbyloopdirectedmutagenesis
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Engineering a bifunctional copper site in the cupredoxin fold by loop-directed mutagenesis

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