Hortensins, Type 1 Ribosome-Inactivating Proteins from Seeds of Red Mountain Spinach: Isolation, Characterization, and Their Effect on Glioblastoma Cells
Ribosome inactivating proteins (RIPs) are specific N-β-glycosylases that are well-characterized in plants. Their enzymatic action is to damage ribosomes, thereby blocking protein translation. Recently, several research groups have been working on the screening for these toxins in edible plants to fa...
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| Format: | Article |
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MDPI AG
2024-03-01
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| Series: | Toxins |
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| Online Access: | https://www.mdpi.com/2072-6651/16/3/135 |
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| author | Sara Ragucci Veronica Russo Angela Clemente Maria Giuseppina Campanile Maria Antonietta Oliva Nicola Landi Paolo Vincenzo Pedone Antonietta Arcella Antimo Di Maro |
| author_facet | Sara Ragucci Veronica Russo Angela Clemente Maria Giuseppina Campanile Maria Antonietta Oliva Nicola Landi Paolo Vincenzo Pedone Antonietta Arcella Antimo Di Maro |
| author_sort | Sara Ragucci |
| collection | DOAJ |
| description | Ribosome inactivating proteins (RIPs) are specific N-β-glycosylases that are well-characterized in plants. Their enzymatic action is to damage ribosomes, thereby blocking protein translation. Recently, several research groups have been working on the screening for these toxins in edible plants to facilitate the use of RIPs as biotechnological tools and biopesticides and to overcome public prejudice. Here, four novel monomeric (type 1) RIPs have been isolated from the seeds of <i>Atriplex hortensis</i> L. var. <i>rubra</i>, which is commonly known as edible red mountain spinach. These enzymes, named hortensins 1, 2, 4, and 5, are able to release the β-fragment and, like many other RIPs, adenines from salmon sperm DNA, thus, acting as polynucleotide:adenosine glycosidases. Structurally, hortensins have a different molecular weight and are purified with different yields (hortensin 1, ~29.5 kDa, 0.28 mg per 100 g; hortensin 2, ~29 kDa, 0.29 mg per 100 g; hortensin 4, ~28.5 kDa, 0.71 mg per 100 g; and hortensin 5, ~30 kDa, 0.65 mg per 100 g); only hortensins 2 and 4 are glycosylated. Furthermore, the major isoforms (hortensins 4 and 5) are cytotoxic toward human continuous glioblastoma U87MG cell line. In addition, the morphological change in U87MG cells in the presence of these toxins is indicative of cell death triggered by the apoptotic pathway, as revealed by nuclear DNA fragmentation (TUNEL assay). |
| first_indexed | 2024-04-24T17:47:47Z |
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| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-04-24T17:47:47Z |
| publishDate | 2024-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-e7a087fef7704ca198f92b27657697e82024-03-27T14:06:22ZengMDPI AGToxins2072-66512024-03-0116313510.3390/toxins16030135Hortensins, Type 1 Ribosome-Inactivating Proteins from Seeds of Red Mountain Spinach: Isolation, Characterization, and Their Effect on Glioblastoma CellsSara Ragucci0Veronica Russo1Angela Clemente2Maria Giuseppina Campanile3Maria Antonietta Oliva4Nicola Landi5Paolo Vincenzo Pedone6Antonietta Arcella7Antimo Di Maro8Department of Environmental, Biological and Pharmaceutical Sciences and Technologies (DiSTABiF), University of Campania ‘Luigi Vanvitelli’, Via Vivaldi 43, 81100 Caserta, ItalyIRCCS Istituto Neurologico Mediterraneo ‘NEUROMED’, Via Atinense 18, 86077 Pozzilli, ItalyDepartment of Environmental, Biological and Pharmaceutical Sciences and Technologies (DiSTABiF), University of Campania ‘Luigi Vanvitelli’, Via Vivaldi 43, 81100 Caserta, ItalyDepartment of Environmental, Biological and Pharmaceutical Sciences and Technologies (DiSTABiF), University of Campania ‘Luigi Vanvitelli’, Via Vivaldi 43, 81100 Caserta, ItalyIRCCS Istituto Neurologico Mediterraneo ‘NEUROMED’, Via Atinense 18, 86077 Pozzilli, ItalyDepartment of Environmental, Biological and Pharmaceutical Sciences and Technologies (DiSTABiF), University of Campania ‘Luigi Vanvitelli’, Via Vivaldi 43, 81100 Caserta, ItalyDepartment of Environmental, Biological and Pharmaceutical Sciences and Technologies (DiSTABiF), University of Campania ‘Luigi Vanvitelli’, Via Vivaldi 43, 81100 Caserta, ItalyIRCCS Istituto Neurologico Mediterraneo ‘NEUROMED’, Via Atinense 18, 86077 Pozzilli, ItalyDepartment of Environmental, Biological and Pharmaceutical Sciences and Technologies (DiSTABiF), University of Campania ‘Luigi Vanvitelli’, Via Vivaldi 43, 81100 Caserta, ItalyRibosome inactivating proteins (RIPs) are specific N-β-glycosylases that are well-characterized in plants. Their enzymatic action is to damage ribosomes, thereby blocking protein translation. Recently, several research groups have been working on the screening for these toxins in edible plants to facilitate the use of RIPs as biotechnological tools and biopesticides and to overcome public prejudice. Here, four novel monomeric (type 1) RIPs have been isolated from the seeds of <i>Atriplex hortensis</i> L. var. <i>rubra</i>, which is commonly known as edible red mountain spinach. These enzymes, named hortensins 1, 2, 4, and 5, are able to release the β-fragment and, like many other RIPs, adenines from salmon sperm DNA, thus, acting as polynucleotide:adenosine glycosidases. Structurally, hortensins have a different molecular weight and are purified with different yields (hortensin 1, ~29.5 kDa, 0.28 mg per 100 g; hortensin 2, ~29 kDa, 0.29 mg per 100 g; hortensin 4, ~28.5 kDa, 0.71 mg per 100 g; and hortensin 5, ~30 kDa, 0.65 mg per 100 g); only hortensins 2 and 4 are glycosylated. Furthermore, the major isoforms (hortensins 4 and 5) are cytotoxic toward human continuous glioblastoma U87MG cell line. In addition, the morphological change in U87MG cells in the presence of these toxins is indicative of cell death triggered by the apoptotic pathway, as revealed by nuclear DNA fragmentation (TUNEL assay).https://www.mdpi.com/2072-6651/16/3/135<i>Atriplex hortensis</i> L.cytotoxicityedible plantsred oracherRNA N-glycosylasesU87MG glioblastoma cells |
| spellingShingle | Sara Ragucci Veronica Russo Angela Clemente Maria Giuseppina Campanile Maria Antonietta Oliva Nicola Landi Paolo Vincenzo Pedone Antonietta Arcella Antimo Di Maro Hortensins, Type 1 Ribosome-Inactivating Proteins from Seeds of Red Mountain Spinach: Isolation, Characterization, and Their Effect on Glioblastoma Cells Toxins <i>Atriplex hortensis</i> L. cytotoxicity edible plants red orache rRNA N-glycosylases U87MG glioblastoma cells |
| title | Hortensins, Type 1 Ribosome-Inactivating Proteins from Seeds of Red Mountain Spinach: Isolation, Characterization, and Their Effect on Glioblastoma Cells |
| title_full | Hortensins, Type 1 Ribosome-Inactivating Proteins from Seeds of Red Mountain Spinach: Isolation, Characterization, and Their Effect on Glioblastoma Cells |
| title_fullStr | Hortensins, Type 1 Ribosome-Inactivating Proteins from Seeds of Red Mountain Spinach: Isolation, Characterization, and Their Effect on Glioblastoma Cells |
| title_full_unstemmed | Hortensins, Type 1 Ribosome-Inactivating Proteins from Seeds of Red Mountain Spinach: Isolation, Characterization, and Their Effect on Glioblastoma Cells |
| title_short | Hortensins, Type 1 Ribosome-Inactivating Proteins from Seeds of Red Mountain Spinach: Isolation, Characterization, and Their Effect on Glioblastoma Cells |
| title_sort | hortensins type 1 ribosome inactivating proteins from seeds of red mountain spinach isolation characterization and their effect on glioblastoma cells |
| topic | <i>Atriplex hortensis</i> L. cytotoxicity edible plants red orache rRNA N-glycosylases U87MG glioblastoma cells |
| url | https://www.mdpi.com/2072-6651/16/3/135 |
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