Interaction between miR4749 and Human Serum Albumin as Revealed by Fluorescence, FRET, Atomic Force Spectroscopy and Computational Modelling
The interaction of Human Serum Albumin (HSA) with the microRNA, miR4749, was investigated by Atomic Force Spectrscopy (AFS), static and time-resolved fluorescence spectroscopy and by computational methods. The formation of a HSA/miR4749 complex with an affinity of about 10<sup>4</sup> M&...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-01-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/3/1291 |
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| author | Valentina Botti Silvia Marrone Salvatore Cannistraro Anna Rita Bizzarri |
| author_facet | Valentina Botti Silvia Marrone Salvatore Cannistraro Anna Rita Bizzarri |
| author_sort | Valentina Botti |
| collection | DOAJ |
| description | The interaction of Human Serum Albumin (HSA) with the microRNA, miR4749, was investigated by Atomic Force Spectrscopy (AFS), static and time-resolved fluorescence spectroscopy and by computational methods. The formation of a HSA/miR4749 complex with an affinity of about 10<sup>4</sup> M<sup>−1</sup> has been assessed through a Stern–Volmer analysis of steady-state fluorescence quenching of the lone Trp residue (Trp214) emission of HSA. Förster Resonance Energy Transfer (FRET) measurements of fluorescence lifetime of the HSA/miR4749 complex were carried out in the absence and in the presence of an acceptor chromophore linked to miR4749. This allowed us to determine a distance of 4.3 ± 0.5 nm between the lone Trp of HSA and the dye bound to miR4749 5p-end. Such a distance was exploited for a screening of the possible binding sites between HSA and miR4749, as predicted by computational docking. Such an approach, further refined by binding free energy calculations, led us to the identification of a consistent model for the structure of the HSA/miR4749 complex in which a positively charged HSA pocket accommodates the negatively charged miRNA molecule. These results designate native HSA as a suitable miRNA carrier under physiological conditions for delivering to appropriate targets. |
| first_indexed | 2024-03-09T23:49:08Z |
| format | Article |
| id | doaj.art-d9775ec5fc074d149c8eda015f875b94 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T23:49:08Z |
| publishDate | 2022-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-d9775ec5fc074d149c8eda015f875b942023-11-23T16:38:08ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-01-01233129110.3390/ijms23031291Interaction between miR4749 and Human Serum Albumin as Revealed by Fluorescence, FRET, Atomic Force Spectroscopy and Computational ModellingValentina Botti0Silvia Marrone1Salvatore Cannistraro2Anna Rita Bizzarri3Biophysics and Nanoscience Centre, Department of Ecology and Biology DEB, Università della Tuscia, Largo dell’Università, 01100 Viterbo, ItalyBiophysics and Nanoscience Centre, Department of Ecology and Biology DEB, Università della Tuscia, Largo dell’Università, 01100 Viterbo, ItalyBiophysics and Nanoscience Centre, Department of Ecology and Biology DEB, Università della Tuscia, Largo dell’Università, 01100 Viterbo, ItalyBiophysics and Nanoscience Centre, Department of Ecology and Biology DEB, Università della Tuscia, Largo dell’Università, 01100 Viterbo, ItalyThe interaction of Human Serum Albumin (HSA) with the microRNA, miR4749, was investigated by Atomic Force Spectrscopy (AFS), static and time-resolved fluorescence spectroscopy and by computational methods. The formation of a HSA/miR4749 complex with an affinity of about 10<sup>4</sup> M<sup>−1</sup> has been assessed through a Stern–Volmer analysis of steady-state fluorescence quenching of the lone Trp residue (Trp214) emission of HSA. Förster Resonance Energy Transfer (FRET) measurements of fluorescence lifetime of the HSA/miR4749 complex were carried out in the absence and in the presence of an acceptor chromophore linked to miR4749. This allowed us to determine a distance of 4.3 ± 0.5 nm between the lone Trp of HSA and the dye bound to miR4749 5p-end. Such a distance was exploited for a screening of the possible binding sites between HSA and miR4749, as predicted by computational docking. Such an approach, further refined by binding free energy calculations, led us to the identification of a consistent model for the structure of the HSA/miR4749 complex in which a positively charged HSA pocket accommodates the negatively charged miRNA molecule. These results designate native HSA as a suitable miRNA carrier under physiological conditions for delivering to appropriate targets.https://www.mdpi.com/1422-0067/23/3/1291miR4749Human Serum Albuminfluorescence quenchingFRETcomputational docking |
| spellingShingle | Valentina Botti Silvia Marrone Salvatore Cannistraro Anna Rita Bizzarri Interaction between miR4749 and Human Serum Albumin as Revealed by Fluorescence, FRET, Atomic Force Spectroscopy and Computational Modelling International Journal of Molecular Sciences miR4749 Human Serum Albumin fluorescence quenching FRET computational docking |
| title | Interaction between miR4749 and Human Serum Albumin as Revealed by Fluorescence, FRET, Atomic Force Spectroscopy and Computational Modelling |
| title_full | Interaction between miR4749 and Human Serum Albumin as Revealed by Fluorescence, FRET, Atomic Force Spectroscopy and Computational Modelling |
| title_fullStr | Interaction between miR4749 and Human Serum Albumin as Revealed by Fluorescence, FRET, Atomic Force Spectroscopy and Computational Modelling |
| title_full_unstemmed | Interaction between miR4749 and Human Serum Albumin as Revealed by Fluorescence, FRET, Atomic Force Spectroscopy and Computational Modelling |
| title_short | Interaction between miR4749 and Human Serum Albumin as Revealed by Fluorescence, FRET, Atomic Force Spectroscopy and Computational Modelling |
| title_sort | interaction between mir4749 and human serum albumin as revealed by fluorescence fret atomic force spectroscopy and computational modelling |
| topic | miR4749 Human Serum Albumin fluorescence quenching FRET computational docking |
| url | https://www.mdpi.com/1422-0067/23/3/1291 |
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