The role of molecular crowding in long-range metalloprotein electron transfer: Dissection into site- and scaffold-specific contributions

Here we report the effect of molecular crowding on long-range protein electron transfer (ET) and disentangle the specific responses of the redox site and the protein milieu. To this end, we studied two different one-electron redox proteins that share the cupredoxin fold but differ in the metal cente...

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Autores principales: Zitare, U.A., Szuster, J., Scocozza, M.F., Espinoza-Cara, A., Leguto, A.J., Morgada, M.N., Vila, A.J., Murgida, D.H.
Formato: JOUR
Materias:
Electron transfer
Frictional control
Loop engineering
Metalloproteins
Molecular crowding
Activation energy
Electron transitions
Friction
Intelligent agents
Metals
Scaffolds
Scaffolds (biology)
Dynamical coupling
Electronic coupling
Metallo-proteins
Metalloprotein
Orthologous proteins
Redox proteins
Proteins
Acceso en línea:http://hdl.handle.net/20.500.12110/paper_00134686_v294_n_p117_Zitare
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Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires
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spelling todo:paper_00134686_v294_n_p117_Zitare2023-10-03T14:10:54Z The role of molecular crowding in long-range metalloprotein electron transfer: Dissection into site- and scaffold-specific contributions Zitare, U.A. Szuster, J. Scocozza, M.F. Espinoza-Cara, A. Leguto, A.J. Morgada, M.N. Vila, A.J. Murgida, D.H. Electron transfer Frictional control Loop engineering Metalloproteins Molecular crowding Activation energy Electron transitions Friction Intelligent agents Metals Scaffolds Scaffolds (biology) Dynamical coupling Electron transfer Electronic coupling Metallo-proteins Metalloprotein Molecular crowding Orthologous proteins Redox proteins Proteins Here we report the effect of molecular crowding on long-range protein electron transfer (ET) and disentangle the specific responses of the redox site and the protein milieu. To this end, we studied two different one-electron redox proteins that share the cupredoxin fold but differ in the metal center, T1 mononuclear blue copper and binuclear CuA, and generated chimeras with hybrid properties by incorporating different T1 centers within the CuA scaffold or by swapping loops between orthologous proteins from different organisms to perturb the CuA site. The heterogeneous ET kinetics of the different proteins was studied by protein film electrochemistry at variable electronic couplings and in the presence of two different crowding agents. The results reveal a strong frictional control of the ET reactions, which for 10 Å tunnelling distances results in a 90% drop of the ET rate when viscosity is matched to that of the mitochondrial interior (ca. 55 cP) by addition of either crowding agent. The effect is ascribed to the dynamical coupling of the metal site and the milieu, which for T1 is found to be twice stronger than for CuA, and the activation energy of protein-solvent motion that is dictated by the overall scaffold. This work highlights the need of explicitly considering molecular crowding effects in protein ET. © 2018 Elsevier Ltd JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_00134686_v294_n_p117_Zitare
institution Universidad de Buenos Aires
institution_str I-28
repository_str R-134
collection Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic Electron transfer
Frictional control
Loop engineering
Metalloproteins
Molecular crowding
Activation energy
Electron transitions
Friction
Intelligent agents
Metals
Scaffolds
Scaffolds (biology)
Dynamical coupling
Electron transfer
Electronic coupling
Metallo-proteins
Metalloprotein
Molecular crowding
Orthologous proteins
Redox proteins
Proteins
spellingShingle Electron transfer
Frictional control
Loop engineering
Metalloproteins
Molecular crowding
Activation energy
Electron transitions
Friction
Intelligent agents
Metals
Scaffolds
Scaffolds (biology)
Dynamical coupling
Electron transfer
Electronic coupling
Metallo-proteins
Metalloprotein
Molecular crowding
Orthologous proteins
Redox proteins
Proteins
Zitare, U.A.
Szuster, J.
Scocozza, M.F.
Espinoza-Cara, A.
Leguto, A.J.
Morgada, M.N.
Vila, A.J.
Murgida, D.H.
The role of molecular crowding in long-range metalloprotein electron transfer: Dissection into site- and scaffold-specific contributions
topic_facet Electron transfer
Frictional control
Loop engineering
Metalloproteins
Molecular crowding
Activation energy
Electron transitions
Friction
Intelligent agents
Metals
Scaffolds
Scaffolds (biology)
Dynamical coupling
Electron transfer
Electronic coupling
Metallo-proteins
Metalloprotein
Molecular crowding
Orthologous proteins
Redox proteins
Proteins
description Here we report the effect of molecular crowding on long-range protein electron transfer (ET) and disentangle the specific responses of the redox site and the protein milieu. To this end, we studied two different one-electron redox proteins that share the cupredoxin fold but differ in the metal center, T1 mononuclear blue copper and binuclear CuA, and generated chimeras with hybrid properties by incorporating different T1 centers within the CuA scaffold or by swapping loops between orthologous proteins from different organisms to perturb the CuA site. The heterogeneous ET kinetics of the different proteins was studied by protein film electrochemistry at variable electronic couplings and in the presence of two different crowding agents. The results reveal a strong frictional control of the ET reactions, which for 10 Å tunnelling distances results in a 90% drop of the ET rate when viscosity is matched to that of the mitochondrial interior (ca. 55 cP) by addition of either crowding agent. The effect is ascribed to the dynamical coupling of the metal site and the milieu, which for T1 is found to be twice stronger than for CuA, and the activation energy of protein-solvent motion that is dictated by the overall scaffold. This work highlights the need of explicitly considering molecular crowding effects in protein ET. © 2018 Elsevier Ltd
format JOUR
author Zitare, U.A.
Szuster, J.
Scocozza, M.F.
Espinoza-Cara, A.
Leguto, A.J.
Morgada, M.N.
Vila, A.J.
Murgida, D.H.
author_facet Zitare, U.A.
Szuster, J.
Scocozza, M.F.
Espinoza-Cara, A.
Leguto, A.J.
Morgada, M.N.
Vila, A.J.
Murgida, D.H.
author_sort Zitare, U.A.
title The role of molecular crowding in long-range metalloprotein electron transfer: Dissection into site- and scaffold-specific contributions
title_short The role of molecular crowding in long-range metalloprotein electron transfer: Dissection into site- and scaffold-specific contributions
title_full The role of molecular crowding in long-range metalloprotein electron transfer: Dissection into site- and scaffold-specific contributions
title_fullStr The role of molecular crowding in long-range metalloprotein electron transfer: Dissection into site- and scaffold-specific contributions
title_full_unstemmed The role of molecular crowding in long-range metalloprotein electron transfer: Dissection into site- and scaffold-specific contributions
title_sort role of molecular crowding in long-range metalloprotein electron transfer: dissection into site- and scaffold-specific contributions
url http://hdl.handle.net/20.500.12110/paper_00134686_v294_n_p117_Zitare
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