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Structural and functional insights into lysine acetylation of cytochrome c using mimetic point mutants
Post‐translational modifications frequently modulate protein functions. Lysine acetylation in particular plays a key role in interactions between respiratory cytochrome c and its metabolic partners. To date, in vivo acetylation of lysines at positions 8 and 53 has specifically been identified in mam...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8634867/ https://www.ncbi.nlm.nih.gov/pubmed/34455704 http://dx.doi.org/10.1002/2211-5463.13284 |
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author | Márquez, Inmaculada Pérez‐Mejías, Gonzalo Guerra‐Castellano, Alejandra Olloqui‐Sariego, José Luis Andreu, Rafael Calvente, Juan José De la Rosa, Miguel A. Díaz‐Moreno, Irene |
author_facet | Márquez, Inmaculada Pérez‐Mejías, Gonzalo Guerra‐Castellano, Alejandra Olloqui‐Sariego, José Luis Andreu, Rafael Calvente, Juan José De la Rosa, Miguel A. Díaz‐Moreno, Irene |
author_sort | Márquez, Inmaculada |
collection | PubMed |
description | Post‐translational modifications frequently modulate protein functions. Lysine acetylation in particular plays a key role in interactions between respiratory cytochrome c and its metabolic partners. To date, in vivo acetylation of lysines at positions 8 and 53 has specifically been identified in mammalian cytochrome c, but little is known about the structural basis of acetylation‐induced functional changes. Here, we independently replaced these two residues in recombinant human cytochrome c with glutamine to mimic lysine acetylation and then characterized the structure and function of the resulting K8Q and K53Q mutants. We found that the physicochemical features were mostly unchanged in the two acetyl‐mimetic mutants, but their thermal stability was significantly altered. NMR chemical shift perturbations of the backbone amide resonances revealed local structural changes, and the thermodynamics and kinetics of electron transfer in mutants immobilized on gold electrodes showed an increase in both protein dynamics and solvent involvement in the redox process. We also observed that the K8Q (but not the K53Q) mutation slightly increased the binding affinity of cytochrome c to its physiological electron donor, cytochrome c (1)—which is a component of mitochondrial complex III, or cytochrome bc (1)—thus suggesting that Lys8 (but not Lys53) is located in the interaction area. Finally, the K8Q and K53Q mutants exhibited reduced efficiency as electron donors to complex IV, or cytochrome c oxidase. |
format | Online Article Text |
id | pubmed-8634867 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-86348672021-12-08 Structural and functional insights into lysine acetylation of cytochrome c using mimetic point mutants Márquez, Inmaculada Pérez‐Mejías, Gonzalo Guerra‐Castellano, Alejandra Olloqui‐Sariego, José Luis Andreu, Rafael Calvente, Juan José De la Rosa, Miguel A. Díaz‐Moreno, Irene FEBS Open Bio Research Articles Post‐translational modifications frequently modulate protein functions. Lysine acetylation in particular plays a key role in interactions between respiratory cytochrome c and its metabolic partners. To date, in vivo acetylation of lysines at positions 8 and 53 has specifically been identified in mammalian cytochrome c, but little is known about the structural basis of acetylation‐induced functional changes. Here, we independently replaced these two residues in recombinant human cytochrome c with glutamine to mimic lysine acetylation and then characterized the structure and function of the resulting K8Q and K53Q mutants. We found that the physicochemical features were mostly unchanged in the two acetyl‐mimetic mutants, but their thermal stability was significantly altered. NMR chemical shift perturbations of the backbone amide resonances revealed local structural changes, and the thermodynamics and kinetics of electron transfer in mutants immobilized on gold electrodes showed an increase in both protein dynamics and solvent involvement in the redox process. We also observed that the K8Q (but not the K53Q) mutation slightly increased the binding affinity of cytochrome c to its physiological electron donor, cytochrome c (1)—which is a component of mitochondrial complex III, or cytochrome bc (1)—thus suggesting that Lys8 (but not Lys53) is located in the interaction area. Finally, the K8Q and K53Q mutants exhibited reduced efficiency as electron donors to complex IV, or cytochrome c oxidase. John Wiley and Sons Inc. 2021-11-09 /pmc/articles/PMC8634867/ /pubmed/34455704 http://dx.doi.org/10.1002/2211-5463.13284 Text en © 2021 The Authors. FEBS Open Bio published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies. https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Articles Márquez, Inmaculada Pérez‐Mejías, Gonzalo Guerra‐Castellano, Alejandra Olloqui‐Sariego, José Luis Andreu, Rafael Calvente, Juan José De la Rosa, Miguel A. Díaz‐Moreno, Irene Structural and functional insights into lysine acetylation of cytochrome c using mimetic point mutants |
title | Structural and functional insights into lysine acetylation of cytochrome c using mimetic point mutants |
title_full | Structural and functional insights into lysine acetylation of cytochrome c using mimetic point mutants |
title_fullStr | Structural and functional insights into lysine acetylation of cytochrome c using mimetic point mutants |
title_full_unstemmed | Structural and functional insights into lysine acetylation of cytochrome c using mimetic point mutants |
title_short | Structural and functional insights into lysine acetylation of cytochrome c using mimetic point mutants |
title_sort | structural and functional insights into lysine acetylation of cytochrome c using mimetic point mutants |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8634867/ https://www.ncbi.nlm.nih.gov/pubmed/34455704 http://dx.doi.org/10.1002/2211-5463.13284 |
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