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Article

Allosteric Communication in the Multifunctional and Redox NQO1 Protein Studied by Cavity-Making Mutations

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Departamento de Química Física, Universidad de Granada, Av. Fuentenueva s/n, 18071 Granada, Spain
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Institute of Microbiology—BioCeV, Academy of Sciences of the Czech Republic, Prumyslova 595, 252 50 Vestec, Czech Republic
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Departamento de Bioquímica y Biología Molecular y Celular, Facultad de Ciencias, Instituto de Biocomputación y Física de Sistemas Complejos (BIFI) (GBsC-CSIC Joint Unit), Universidad de Zaragoza, 50009 Zaragoza, Spain
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Institute of Biotechnology—BioCeV, Academy of Sciences of the Czech Republic, Prumyslova 595, 252 50 Vestec, Czech Republic
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Department of Biochemistry, Faculty of Science, Charles University, Hlavova 2030/8, 128 43 Prague, Czech Republic
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Departamento de Bioquímica y Biología Molecular I, Facultad de Ciencias y Centro de Investigaciones Biomédicas (CIBM), Universidad de Granada, 18016 Granada, Spain
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Center for Rare Diseases (CIBERER), Hospital Universitario de Canarias, Universidad de la Laguna, 38320 Tenerife, Spain
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Department of Biotechnology, Bhupat & Jyoti Mehta School of Biosciences, Indian Institute of Technology Madras (IITM), Chennai 600036, India
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Departamento de Química Física, Unidad de Excelencia en Química Aplicada a Biomedicina y Medioambiente e Instituto de Biotecnología, Universidad de Granada, Av. Fuentenueva s/n, 18071 Granada, Spain
*
Authors to whom correspondence should be addressed.
These authors contributed equally to this work.
Academic Editor: Alessandra Napolitano
Antioxidants 2022, 11(6), 1110; https://doi.org/10.3390/antiox11061110
Received: 26 April 2022 / Revised: 28 May 2022 / Accepted: 30 May 2022 / Published: 2 June 2022
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. View Full-Text
Keywords: antioxidant defense; flavoprotein; FAD binding; structural perturbation; protein core; allosterism; cavity-making mutation antioxidant defense; flavoprotein; FAD binding; structural perturbation; protein core; allosterism; cavity-making mutation
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MDPI and ACS Style

Pacheco-Garcia, J.L.; Loginov, D.S.; Anoz-Carbonell, E.; Vankova, P.; Palomino-Morales, R.; Salido, E.; Man, P.; Medina, M.; Naganathan, A.N.; Pey, A.L. Allosteric Communication in the Multifunctional and Redox NQO1 Protein Studied by Cavity-Making Mutations. Antioxidants 2022, 11, 1110. https://doi.org/10.3390/antiox11061110

AMA Style

Pacheco-Garcia JL, Loginov DS, Anoz-Carbonell E, Vankova P, Palomino-Morales R, Salido E, Man P, Medina M, Naganathan AN, Pey AL. Allosteric Communication in the Multifunctional and Redox NQO1 Protein Studied by Cavity-Making Mutations. Antioxidants. 2022; 11(6):1110. https://doi.org/10.3390/antiox11061110

Chicago/Turabian Style

Pacheco-Garcia, Juan L., Dmitry S. Loginov, Ernesto Anoz-Carbonell, Pavla Vankova, Rogelio Palomino-Morales, Eduardo Salido, Petr Man, Milagros Medina, Athi N. Naganathan, and Angel L. Pey. 2022. "Allosteric Communication in the Multifunctional and Redox NQO1 Protein Studied by Cavity-Making Mutations" Antioxidants 11, no. 6: 1110. https://doi.org/10.3390/antiox11061110

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