V.О. Shlyakhovenko1, І.І. Ganusevich1, О.А. Samoylenko1, Yu.M. Samchenko2, А.V. Verbinenko1, O.A Solovyova2

1RE Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology, NAS of Ukraine,

2FD Ovcharenko Institute of Biocolloidal Chemistry, NAS of Ukraine, Kyiv, Ukraine


Summary. Aim: to investigate the possibility of enzymatic reactivation of RNase activity of peripheral blood cells of patients with colorectal cancer (CRC) after sorption on nanoplates Laponite® RD (Lap). Objects and methods: the study was performed with the cell suspension of peripheral blood of CRC patients. Samples of cell lysates were combined with a 1% suspension of Lap nanoplates. Then RNase was extracted with 0,25 N H2SO4 or 2% solution of sodium dodecyl sulfate (DDS). The zymogram technique was used to analyze RNase activity. Results: it was found that RNases bind with nanoplates Lap and form complexes with loss of enzymatic activity. It is known that RNase can be released from the complex by extraction with 0,25 N H2SO4 or 2% sodium DDS solution. RNase is able to restore its enzymatic activity when extraction from the complex with a 2% sodium DDS solution is used. But with the extraction of 0,25 N H2SO4, the enzymatic activity is irreversibly lost. Conclusion: RNase extracted from the nanoplates Lap can be active again as an enzyme that catalyzes the cleavage of RNA and hybrid RNA/DNA molecules, depending on the method of extraction.

Keywords: Lap nanoplates, RNase reactivation, zymograms.



1. Peixoto D, Pereira I, Pereira-Silva M, et al. Emerging role of nanoclays in cancer research, diagnosis, and therapy. Coord Chem Rev 2021; 440: 213956.

2. Lee VT, Sondermann H, Winkler WC. Nano-RNases: oligo-or dinucleases? FEMS Microbiol Rev 2022; 46: fuac038. doi: 10.1093/femsre/fuac038.

3. Wang Y, Abrol R, Mak JYW, et al. Histone deacetylase 7: a signalling hub controlling development, inflammation, metabolism and disease. FEBS J 2023; 290: 2805–32. doi: 10.1111/febs.16437.

4. Stealey ST, Gaharwar AK, Zustiak SP. Laponite-based nanocomposite hydrogels for drug delivery applications. Pharmaceuticals 2023; 16 (6): 821.

5. Eller CH, Raines RT. Antimicrobial synergy of a ribonuclease and a peptide secreted by human cells. ACS Infect Dis 2020; 6 (11): 3083–8. doi: 10.1021/acsinfecdis.0c00594.

6. Artman M, Fry M, Engelberg H. The preparation and characterization of ribonucleic acid obtained by direct phenol extraction of intact cells of Escherichia coli from low ionic environment. Biochem Biophys Res Commun 1966; 25 (1): 49–53. doi: 10.1016/0006-291x(66)90638-3.

7. Samoylenko O, Korotych O, Manilo M, et al. Biomedical applications of laponite{\textregistered}-based nanomaterials and formulations. In: Soft Matter Systems for Biomedical Applications. Bulavin L, Lebovka N (eds). Springer Nature, 2021, 385–452. doi:10.1007/978-3-030-80924-9_15.

8. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227 (5259): 680–5. doi: 10.1038/227680a0.

9. Shlyakhovenko V, Samoylenko O. Photopolymerization with EDTA and riboflavin for proteins analysis in polyacrylamide gel electrophoresis. Protein J 2022; 41 (4–5): 438–43. doi: 10.1007/s10930-022-10068-3.

10. Ohgi K, Sand A, Takizawa Y, Jrie M. Purification of acid ribonucleases from bovine spleen. J Biochem 1988; 103 (2): 267–73. doi: 10.1093/oxfordjournals.jbchem.a122259.

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