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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MDPI AG
2021-02-01
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| Series: | Life |
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| Online Access: | https://www.mdpi.com/2075-1729/11/2/168 |
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| author | Riccardo Montioli Rachele Campagnari Sabrina Fasoli Andrea Fagagnini Andra Caloiu Marcello Smania Marta Menegazzi Giovanni Gotte |
| author_facet | Riccardo Montioli Rachele Campagnari Sabrina Fasoli Andrea Fagagnini Andra Caloiu Marcello Smania Marta Menegazzi Giovanni Gotte |
| author_sort | Riccardo Montioli |
| collection | DOAJ |
| 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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| format | Article |
| id | doaj.art-728fafd679174371a2d75da9c25bd661 |
| institution | Directory Open Access Journal |
| issn | 2075-1729 |
| language | English |
| last_indexed | 2024-03-09T00:40:40Z |
| publishDate | 2021-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Life |
| spelling | doaj.art-728fafd679174371a2d75da9c25bd6612023-12-11T17:52:56ZengMDPI AGLife2075-17292021-02-0111216810.3390/life11020168RNase A Domain-Swapped Dimers Produced Through Different Methods: Structure–Catalytic Properties and Antitumor ActivityRiccardo Montioli0Rachele Campagnari1Sabrina Fasoli2Andrea Fagagnini3Andra Caloiu4Marcello Smania5Marta Menegazzi6Giovanni Gotte7Department of Neuroscience, Biomedicine, and Movement Sciences, Biological Chemistry Section, University of Verona, Strada Le Grazie, 8, I-37134 Verona, ItalyDepartment of Neuroscience, Biomedicine, and Movement Sciences, Biological Chemistry Section, University of Verona, Strada Le Grazie, 8, I-37134 Verona, ItalyDepartment of Neuroscience, Biomedicine, and Movement Sciences, Biological Chemistry Section, University of Verona, Strada Le Grazie, 8, I-37134 Verona, ItalyDepartment of Neuroscience, Biomedicine, and Movement Sciences, Biological Chemistry Section, University of Verona, Strada Le Grazie, 8, I-37134 Verona, ItalyDepartment of Microbiology and Virology, Wexham Park Hospital, Wexham Road, Slough SL24HL, Berkshire, UKDepartment of Neuroscience, Biomedicine, and Movement Sciences, Biological Chemistry Section, University of Verona, Strada Le Grazie, 8, I-37134 Verona, ItalyDepartment of Neuroscience, Biomedicine, and Movement Sciences, Biological Chemistry Section, University of Verona, Strada Le Grazie, 8, I-37134 Verona, ItalyDepartment of Neuroscience, Biomedicine, and Movement Sciences, Biological Chemistry Section, University of Verona, Strada Le Grazie, 8, I-37134 Verona, ItalyUpon 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.https://www.mdpi.com/2075-1729/11/2/168RNase ARNase A dimers3D domain swappingenzymatic activitycytotoxic activitymelanoma cells |
| spellingShingle | Riccardo Montioli Rachele Campagnari Sabrina Fasoli Andrea Fagagnini Andra Caloiu Marcello Smania Marta Menegazzi Giovanni Gotte RNase A Domain-Swapped Dimers Produced Through Different Methods: Structure–Catalytic Properties and Antitumor Activity Life RNase A RNase A dimers 3D domain swapping enzymatic activity cytotoxic activity melanoma cells |
| 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 | RNase A RNase A dimers 3D domain swapping enzymatic activity cytotoxic activity melanoma cells |
| url | https://www.mdpi.com/2075-1729/11/2/168 |
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