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Allosteric Communication in the Multifunctional and Redox NQO1 Protein Studied by Cavity-Making Mutations
Allosterism is a common phenomenon in protein biochemistry that allows rapid regulation of protein stability; dynamics and function. However, the mechanisms by which allosterism occurs (by mutations or post-translational modifications (PTMs)) may be complex, particularly due to long-range propagatio...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9219786/ https://www.ncbi.nlm.nih.gov/pubmed/35740007 http://dx.doi.org/10.3390/antiox11061110 |
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author | Pacheco-Garcia, Juan Luis Loginov, Dmitry S. Anoz-Carbonell, Ernesto Vankova, Pavla Palomino-Morales, Rogelio Salido, Eduardo Man, Petr Medina, Milagros Naganathan, Athi N. Pey, Angel L. |
author_facet | Pacheco-Garcia, Juan Luis Loginov, Dmitry S. Anoz-Carbonell, Ernesto Vankova, Pavla Palomino-Morales, Rogelio Salido, Eduardo Man, Petr Medina, Milagros Naganathan, Athi N. Pey, Angel L. |
author_sort | Pacheco-Garcia, Juan Luis |
collection | PubMed |
description | Allosterism is a common phenomenon in protein biochemistry that allows rapid regulation of protein stability; dynamics and function. However, the mechanisms by which allosterism occurs (by mutations or post-translational modifications (PTMs)) may be complex, particularly due to long-range propagation of the perturbation across protein structures. In this work, we have investigated allosteric communication in the multifunctional, cancer-related and antioxidant protein NQO1 by mutating several fully buried leucine residues (L7, L10 and L30) to smaller residues (V, A and G) at sites in the N-terminal domain. In almost all cases, mutated residues were not close to the FAD or the active site. Mutations L→G strongly compromised conformational stability and solubility, and L30A and L30V also notably decreased solubility. The mutation L10A, closer to the FAD binding site, severely decreased FAD binding affinity (≈20 fold vs. WT) through long-range and context-dependent effects. Using a combination of experimental and computational analyses, we show that most of the effects are found in the apo state of the protein, in contrast to other common polymorphisms and PTMs previously characterized in NQO1. The integrated study presented here is a first step towards a detailed structural–functional mapping of the mutational landscape of NQO1, a multifunctional and redox signaling protein of high biomedical relevance. |
format | Online Article Text |
id | pubmed-9219786 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-92197862022-06-24 Allosteric Communication in the Multifunctional and Redox NQO1 Protein Studied by Cavity-Making Mutations Pacheco-Garcia, Juan Luis Loginov, Dmitry S. Anoz-Carbonell, Ernesto Vankova, Pavla Palomino-Morales, Rogelio Salido, Eduardo Man, Petr Medina, Milagros Naganathan, Athi N. Pey, Angel L. Antioxidants (Basel) Article Allosterism is a common phenomenon in protein biochemistry that allows rapid regulation of protein stability; dynamics and function. However, the mechanisms by which allosterism occurs (by mutations or post-translational modifications (PTMs)) may be complex, particularly due to long-range propagation of the perturbation across protein structures. In this work, we have investigated allosteric communication in the multifunctional, cancer-related and antioxidant protein NQO1 by mutating several fully buried leucine residues (L7, L10 and L30) to smaller residues (V, A and G) at sites in the N-terminal domain. In almost all cases, mutated residues were not close to the FAD or the active site. Mutations L→G strongly compromised conformational stability and solubility, and L30A and L30V also notably decreased solubility. The mutation L10A, closer to the FAD binding site, severely decreased FAD binding affinity (≈20 fold vs. WT) through long-range and context-dependent effects. Using a combination of experimental and computational analyses, we show that most of the effects are found in the apo state of the protein, in contrast to other common polymorphisms and PTMs previously characterized in NQO1. The integrated study presented here is a first step towards a detailed structural–functional mapping of the mutational landscape of NQO1, a multifunctional and redox signaling protein of high biomedical relevance. MDPI 2022-06-02 /pmc/articles/PMC9219786/ /pubmed/35740007 http://dx.doi.org/10.3390/antiox11061110 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Pacheco-Garcia, Juan Luis Loginov, Dmitry S. Anoz-Carbonell, Ernesto Vankova, Pavla Palomino-Morales, Rogelio Salido, Eduardo Man, Petr Medina, Milagros Naganathan, Athi N. Pey, Angel L. Allosteric Communication in the Multifunctional and Redox NQO1 Protein Studied by Cavity-Making Mutations |
title | Allosteric Communication in the Multifunctional and Redox NQO1 Protein Studied by Cavity-Making Mutations |
title_full | Allosteric Communication in the Multifunctional and Redox NQO1 Protein Studied by Cavity-Making Mutations |
title_fullStr | Allosteric Communication in the Multifunctional and Redox NQO1 Protein Studied by Cavity-Making Mutations |
title_full_unstemmed | Allosteric Communication in the Multifunctional and Redox NQO1 Protein Studied by Cavity-Making Mutations |
title_short | Allosteric Communication in the Multifunctional and Redox NQO1 Protein Studied by Cavity-Making Mutations |
title_sort | allosteric communication in the multifunctional and redox nqo1 protein studied by cavity-making mutations |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9219786/ https://www.ncbi.nlm.nih.gov/pubmed/35740007 http://dx.doi.org/10.3390/antiox11061110 |
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