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RNase A Domain-Swapped Dimers Produced Through Different Methods: Structure–Catalytic Properties and Antitumor Activity

Upon oligomerization, RNase A can acquire important properties, such as cytotoxicity against leukemic cells. When lyophilized from 40% acetic acid solutions, the enzyme self-associates through the so-called three-dimensional domain swapping (3D-DS) mechanism involving both N- and/or C-terminals. The...

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Autores principales: Montioli, Riccardo, Campagnari, Rachele, Fasoli, Sabrina, Fagagnini, Andrea, Caloiu, Andra, Smania, Marcello, Menegazzi, Marta, Gotte, Giovanni
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7926746/
https://www.ncbi.nlm.nih.gov/pubmed/33669993
http://dx.doi.org/10.3390/life11020168
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author Montioli, Riccardo
Campagnari, Rachele
Fasoli, Sabrina
Fagagnini, Andrea
Caloiu, Andra
Smania, Marcello
Menegazzi, Marta
Gotte, Giovanni
author_facet Montioli, Riccardo
Campagnari, Rachele
Fasoli, Sabrina
Fagagnini, Andrea
Caloiu, Andra
Smania, Marcello
Menegazzi, Marta
Gotte, Giovanni
author_sort Montioli, Riccardo
collection PubMed
description Upon oligomerization, RNase A can acquire important properties, such as cytotoxicity against leukemic cells. When lyophilized from 40% acetic acid solutions, the enzyme self-associates through the so-called three-dimensional domain swapping (3D-DS) mechanism involving both N- and/or C-terminals. The same species are formed if the enzyme is subjected to thermal incubation in various solvents, especially in 40% ethanol. We evaluated here if significant structural modifications might occur in RNase A N- or C-swapped dimers and/or in the residual monomer(s), as a function of the oligomerization protocol applied. We detected that the monomer activity vs. ss-RNA was partly affected by both protocols, although the protein does not suffer spectroscopic alterations. Instead, the two N-swapped dimers showed differences in the fluorescence emission spectra but almost identical enzymatic activities, while the C-swapped dimers displayed slightly different activities vs. both ss- or ds-RNA substrates together with not negligible fluorescence emission alterations within each other. Besides these results, we also discuss the reasons justifying the different relative enzymatic activities displayed by the N-dimers and C-dimers. Last, similarly with data previously registered in a mouse model, we found that both dimeric species significantly decrease human melanoma A375 cell viability, while only N-dimers reduce human melanoma MeWo cell growth.
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spelling pubmed-79267462021-03-04 RNase A Domain-Swapped Dimers Produced Through Different Methods: Structure–Catalytic Properties and Antitumor Activity Montioli, Riccardo Campagnari, Rachele Fasoli, Sabrina Fagagnini, Andrea Caloiu, Andra Smania, Marcello Menegazzi, Marta Gotte, Giovanni Life (Basel) Article Upon oligomerization, RNase A can acquire important properties, such as cytotoxicity against leukemic cells. When lyophilized from 40% acetic acid solutions, the enzyme self-associates through the so-called three-dimensional domain swapping (3D-DS) mechanism involving both N- and/or C-terminals. The same species are formed if the enzyme is subjected to thermal incubation in various solvents, especially in 40% ethanol. We evaluated here if significant structural modifications might occur in RNase A N- or C-swapped dimers and/or in the residual monomer(s), as a function of the oligomerization protocol applied. We detected that the monomer activity vs. ss-RNA was partly affected by both protocols, although the protein does not suffer spectroscopic alterations. Instead, the two N-swapped dimers showed differences in the fluorescence emission spectra but almost identical enzymatic activities, while the C-swapped dimers displayed slightly different activities vs. both ss- or ds-RNA substrates together with not negligible fluorescence emission alterations within each other. Besides these results, we also discuss the reasons justifying the different relative enzymatic activities displayed by the N-dimers and C-dimers. Last, similarly with data previously registered in a mouse model, we found that both dimeric species significantly decrease human melanoma A375 cell viability, while only N-dimers reduce human melanoma MeWo cell growth. MDPI 2021-02-21 /pmc/articles/PMC7926746/ /pubmed/33669993 http://dx.doi.org/10.3390/life11020168 Text en © 2021 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
Montioli, Riccardo
Campagnari, Rachele
Fasoli, Sabrina
Fagagnini, Andrea
Caloiu, Andra
Smania, Marcello
Menegazzi, Marta
Gotte, Giovanni
RNase A Domain-Swapped Dimers Produced Through Different Methods: Structure–Catalytic Properties and Antitumor Activity
title RNase A Domain-Swapped Dimers Produced Through Different Methods: Structure–Catalytic Properties and Antitumor Activity
title_full RNase A Domain-Swapped Dimers Produced Through Different Methods: Structure–Catalytic Properties and Antitumor Activity
title_fullStr RNase A Domain-Swapped Dimers Produced Through Different Methods: Structure–Catalytic Properties and Antitumor Activity
title_full_unstemmed RNase A Domain-Swapped Dimers Produced Through Different Methods: Structure–Catalytic Properties and Antitumor Activity
title_short RNase A Domain-Swapped Dimers Produced Through Different Methods: Structure–Catalytic Properties and Antitumor Activity
title_sort rnase a domain-swapped dimers produced through different methods: structure–catalytic properties and antitumor activity
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7926746/
https://www.ncbi.nlm.nih.gov/pubmed/33669993
http://dx.doi.org/10.3390/life11020168
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