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The Role of a Key Amino Acid Position in Species-Specific Proteinaceous dUTPase Inhibition
Protein inhibitors of key DNA repair enzymes play an important role in deciphering physiological pathways responsible for genome integrity, and may also be exploited in biomedical research. The staphylococcal repressor StlSaPIbov1 protein was described to be an efficient inhibitor of dUTPase homolog...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6627510/ https://www.ncbi.nlm.nih.gov/pubmed/31174420 http://dx.doi.org/10.3390/biom9060221 |
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author | Benedek, András Temesváry-Kis, Fanni Khatanbaatar, Tamjidmaa Leveles, Ibolya Surányi, Éva Viola Szabó, Judit Eszter Wunderlich, Lívius Vértessy, Beáta G. |
author_facet | Benedek, András Temesváry-Kis, Fanni Khatanbaatar, Tamjidmaa Leveles, Ibolya Surányi, Éva Viola Szabó, Judit Eszter Wunderlich, Lívius Vértessy, Beáta G. |
author_sort | Benedek, András |
collection | PubMed |
description | Protein inhibitors of key DNA repair enzymes play an important role in deciphering physiological pathways responsible for genome integrity, and may also be exploited in biomedical research. The staphylococcal repressor StlSaPIbov1 protein was described to be an efficient inhibitor of dUTPase homologues showing a certain degree of species-specificity. In order to provide insight into the inhibition mechanism, in the present study we investigated the interaction of StlSaPIbov1 and Escherichia coli dUTPase. Although we observed a strong interaction of these proteins, unexpectedly the E. coli dUTPase was not inhibited. Seeking a structural explanation for this phenomenon, we identified a key amino acid position where specific mutations sensitized E. coli dUTPase to StlSaPIbov1 inhibition. We solved the three-dimensional (3D) crystal structure of such a mutant in complex with the substrate analogue dUPNPP and surprisingly found that the C-terminal arm of the enzyme, containing the P-loop-like motif was ordered in the structure. This segment was never localized before in any other E. coli dUTPase crystal structures. The 3D structure in agreement with solution phase experiments suggested that ordering of the flexible C-terminal segment upon substrate binding is a major factor in defining the sensitivity of E. coli dUTPase for StlSaPIbov1 inhibition. |
format | Online Article Text |
id | pubmed-6627510 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-66275102019-07-23 The Role of a Key Amino Acid Position in Species-Specific Proteinaceous dUTPase Inhibition Benedek, András Temesváry-Kis, Fanni Khatanbaatar, Tamjidmaa Leveles, Ibolya Surányi, Éva Viola Szabó, Judit Eszter Wunderlich, Lívius Vértessy, Beáta G. Biomolecules Article Protein inhibitors of key DNA repair enzymes play an important role in deciphering physiological pathways responsible for genome integrity, and may also be exploited in biomedical research. The staphylococcal repressor StlSaPIbov1 protein was described to be an efficient inhibitor of dUTPase homologues showing a certain degree of species-specificity. In order to provide insight into the inhibition mechanism, in the present study we investigated the interaction of StlSaPIbov1 and Escherichia coli dUTPase. Although we observed a strong interaction of these proteins, unexpectedly the E. coli dUTPase was not inhibited. Seeking a structural explanation for this phenomenon, we identified a key amino acid position where specific mutations sensitized E. coli dUTPase to StlSaPIbov1 inhibition. We solved the three-dimensional (3D) crystal structure of such a mutant in complex with the substrate analogue dUPNPP and surprisingly found that the C-terminal arm of the enzyme, containing the P-loop-like motif was ordered in the structure. This segment was never localized before in any other E. coli dUTPase crystal structures. The 3D structure in agreement with solution phase experiments suggested that ordering of the flexible C-terminal segment upon substrate binding is a major factor in defining the sensitivity of E. coli dUTPase for StlSaPIbov1 inhibition. MDPI 2019-06-06 /pmc/articles/PMC6627510/ /pubmed/31174420 http://dx.doi.org/10.3390/biom9060221 Text en © 2019 by the authors. 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 (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Benedek, András Temesváry-Kis, Fanni Khatanbaatar, Tamjidmaa Leveles, Ibolya Surányi, Éva Viola Szabó, Judit Eszter Wunderlich, Lívius Vértessy, Beáta G. The Role of a Key Amino Acid Position in Species-Specific Proteinaceous dUTPase Inhibition |
title | The Role of a Key Amino Acid Position in Species-Specific Proteinaceous dUTPase Inhibition |
title_full | The Role of a Key Amino Acid Position in Species-Specific Proteinaceous dUTPase Inhibition |
title_fullStr | The Role of a Key Amino Acid Position in Species-Specific Proteinaceous dUTPase Inhibition |
title_full_unstemmed | The Role of a Key Amino Acid Position in Species-Specific Proteinaceous dUTPase Inhibition |
title_short | The Role of a Key Amino Acid Position in Species-Specific Proteinaceous dUTPase Inhibition |
title_sort | role of a key amino acid position in species-specific proteinaceous dutpase inhibition |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6627510/ https://www.ncbi.nlm.nih.gov/pubmed/31174420 http://dx.doi.org/10.3390/biom9060221 |
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