Aqueous self-assembly of extracted cyclotides from Viola odorata into novel stable supramolecular structures

• Published in: Journal of molecular liquids 2023-02, Vol.372, p.121217, Article 121217
• Main Authors: Dayani, Ladan, Hashempour, Hossein, Varshosaz, Jaleh, Aliomrani, Mehdi, Sadeghi Dinani, Masoud, Taheri, Azade
• Format: Article
• Language: English
• Subjects: Cyclotides; Peptide nanotubes; Self-assembly; Stability; Viola odorata
• ISSN: 0167-7322; 1873-3166
• DOI: 10.1016/j.molliq.2023.121217

[Display omitted] •Cyclotides have the potential ability to be self-assembled into nanostructures in different media.•The pH, temperature, time and peptide concentration have effect on size, PDI, zeta potential of final nanostructures.•Self-assembled cyclotides showed desirable stability in different conditions.•Self-assembled cyclotides could be used for different application such as drug delivery. Cyclotides are categorized as cyclic peptides consisting of 28–37 amino acids. The cyclic nature, as well as the presence of three intramolecular disulfide bonds, resulted in the extraordinary stability of the cyclotides. In addition, they have the ability to self-associate. In this study, we investigated the ability of the cyclotides to be self-assembled by UV–visible spectroscopy. Then after, the isoelectric points of cyclotides were determined by potentiometer titration and the zeta potential method. Also, the effects of time (1, 2, 3, 5, 7, 10 days), temperature (25 °C, 37 °C), pH (2, 4, 6), and peptide concentration (100, 1000, 5000 µg/ml) on size, poly dispersity index (PDI) and zeta potential of the self-assembled nanostructures were investigated. The stability of the cyclotide nanotubes after freeze-drying, 50-fold dilution with phosphate buffered saline (PBS), and exposure to the fetal bovine serum (FBS) were studied too. The results demonstrated that the cyclotides can self-assemble in different studied media. The potentiometer titration and zeta potential measurements confirmed that the isoelectric points of cyclotides were near the pH of 4. From day 0 to 10, the size of nanostructures increased and at pH 4, the largest nanostructures were obtained at day 10, which is due to the fact that the isoelectric points of the cyclotides were near this pH. Also, increasing the cyclotide concentration increased the size of the assemblies. Furthermore, the effect of temperature is largely depending on pH in which it seems that at pH 2 and 37 °C, the aggregation of cyclotides were facilitated than 25 °C. An interesting finding was that the cyclotide nanotubes showed exceptional stability after freeze-drying, exposure to FBS, and dilution by PBS, suggesting that the prepared cyclotide nanotubes are excellent candidates to be used in biological fields, for example, in drug delivery purposes.