Kirkiin: A New Toxic Type 2 Ribosome-Inactivating Protein from the Caudex of Adenia kirkii
Abstract
:1. Introduction
2. Results
2.1. Purification and Characterization of Adenia Kirkii Lectins
2.2. Enzymatic Properties of Kirkiin
2.2.1. Effect on Protein Synthesis
2.2.2. rRNA N-Glycosylase Activity on Mammalian and Yeast Ribosomes
2.2.3. DNA and RNA Adenine Polynucleotide Glycosylase Activity
2.2.4. Endonuclease Activity on Supercoiled Plasmid DNA
2.3. Immunological Properties
2.4. Cytotoxic Effects
2.4.1. Effect of Kirkiin on NB100 Protein Synthesis and Cell Viability
2.4.2. Evaluation of Apoptosis Induced by Kirkiin in NB100 Cells
3. Discussion
4. Conclusions
5. Materials and Methods
5.1. Materials
5.2. Methods
5.2.1. Adenia Kirkii Lectin Purification
5.2.2. Cell Free Protein Synthesis Inhibition
5.2.3. Hemagglutinating Activity
5.2.4. rRNA Glycosylase Activity
5.2.5. Adenine Polynucleotide Glycosylase Activity on Salmon Sperm DNA and on Tobacco Mosaic Virus (TMV) RNA
5.2.6. Endonuclease Activity on Supercoiled Plasmid DNA
5.2.7. Enzyme-Linked Immunosorbent Assay (ELISA)
5.2.8. Cell Protein Synthesis Inhibition and Viability Assay
5.2.9. Evaluation of Apoptosis
5.2.10. Statistical Analyses
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Adenia Species | Cell Free IC50 (μg/mL) | Agglutinating Activity 1 (μg/mL) | Ref. | |
---|---|---|---|---|
Non-Red. | Red. | |||
A. kirkii | 9.2 | 4.7 | 4.0 | |
A. stenodactyla | 5.6 | 0.5 | 49.9 | [17] |
A. lanceolate | 5.2 | 1.1 | 230.9 | [17] |
A. volkensii | 7.5 | 0.7 | 15.6 | [15,17] |
Purification Step | Protein (mg/mL) | Total Protein (mg) | Total Protein (%) | IC50 (µg/mL) 1 | Agglutinating Activity 2 (μg/mL) | Specific Activity (U/mg) 3 | Total Activity (U) | Yeld (%) | |
---|---|---|---|---|---|---|---|---|---|
Non-Red. | Red. | ||||||||
Crude extract | 3.46 | 1730.0 | 100.0 | 55.0 | 21.2 | 45.1 | 18.2 | 31,454.5 | 100.0 |
Sepharose CL-6B Eluate | 2.84 | 107.9 | 6.2 | 9.2 | 4.7 | 4.0 | 108.7 | 4130.4 | 13.1 |
Sephacryl S-100 peak 1 | 1.43 | 24.2 | 1.4 | 7.4 | 1.0 | 175.0 | 135.1 | 3270.2 | 10.4 |
Sephacryl S-100 peak 2 | 1.49 | 62.4 | 3.6 | >50 | >50 | 2.9 | - | - | - |
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Bortolotti, M.; Maiello, S.; Ferreras, J.M.; Iglesias, R.; Polito, L.; Bolognesi, A. Kirkiin: A New Toxic Type 2 Ribosome-Inactivating Protein from the Caudex of Adenia kirkii. Toxins 2021, 13, 81. https://doi.org/10.3390/toxins13020081
Bortolotti M, Maiello S, Ferreras JM, Iglesias R, Polito L, Bolognesi A. Kirkiin: A New Toxic Type 2 Ribosome-Inactivating Protein from the Caudex of Adenia kirkii. Toxins. 2021; 13(2):81. https://doi.org/10.3390/toxins13020081
Chicago/Turabian StyleBortolotti, Massimo, Stefania Maiello, José M. Ferreras, Rosario Iglesias, Letizia Polito, and Andrea Bolognesi. 2021. "Kirkiin: A New Toxic Type 2 Ribosome-Inactivating Protein from the Caudex of Adenia kirkii" Toxins 13, no. 2: 81. https://doi.org/10.3390/toxins13020081
APA StyleBortolotti, M., Maiello, S., Ferreras, J. M., Iglesias, R., Polito, L., & Bolognesi, A. (2021). Kirkiin: A New Toxic Type 2 Ribosome-Inactivating Protein from the Caudex of Adenia kirkii. Toxins, 13(2), 81. https://doi.org/10.3390/toxins13020081