Spectrophotometric investigation of the usefulness of novel pyridine derivatives to the role of luminescent sensors for the determination of albumin
Authors:
- Patryk Szymaszek,
- Joanna Ortyl
Abstract
Proteins are considered one of the most critical bioactive molecules due to their involvement in nutritional, metabolic and immune processes. The content of biomacromolecules in body fluid can be used in diagnostics as a vital indicator. Therefore, the determination of blood proteins is essential in medical diagnosis. Albumins are the most abundant proteins in the body fluid, playing an essential role in transporting various ligands such as ions, lipids, drugs, toxins. Bovine serum albumin (BSA) is widely used as a model protein to evaluate the interaction of active compounds with albumin due to its low cost, good availability and structural homology with human serum albumin (HAS). Structurally, BSA is a large globular protein (66.4 kDa), consisting of 538 amino acids arranged in a single-chain polypeptide. BSA shares 76% of the sequence with HAS and contains two tryptophan (Try) amino acid residues, Trp-134 and Trp-212, located in domains I and II, respectively. The functional properties of 2-amino-4,6-diphenyl-pyridine-3-carbonitrile derivatives for albumin determination were performed by fluorescence emission of Trp residues and fluorescence of protein-bound sensors. In the wide range of methods for determining drug-protein interactions, fluorescence techniques are the most popular due to their high sensitivity, speed and ease of execution. This study aimed to investigate the interaction and affinity of novel pyridine derivatives to BSA by UV-Vis spectroscopy and fluorescence spectroscopy to determine the feasibility of using the tested derivatives as fluorescent sensors for quantitative and qualitative determination of albumin by determining the binding parameters such as Stern-Volmer constant, binding constant, number of binding sites, binding location and thermodynamic parameters of binding. Additionally, FRET experiment and temperature-dependent emission experiment were carried out to determine the type of quenching mechanism of fluorescence. Based on the studies, it can be concluded that the sensor-BSA interaction leads to increase in emission intensity. The calculated binding parameters allowed us to conclude that the investigated sensors bind to BSA in 1:1 stoichiometry with moderate strength, and the quenching for T1 and T3 probes results from static interaction. The calculated negative value of ΔG indicates that the complexation is a spontaneous process. Moreover, based on the estimated values of entropy and enthalpy change, it was determined that the binding of T1 and T3 sensors to BSA occurred mainly due to van der Waals and hydrogen bonding interactions. The determined values of LOD and LOQ are on the order of magnitude of 10-7 M. The Förster experiment indicate a high probability of the occurrence of energy transfer. The binding site marker experiment showed that all probes tested bind to the site II of BSA. Based on the determined parameters, it can be concluded that the tested probes are suitable for quantitative and qualitative determination of albumin.
- Record ID
- CUT7f4859bc081a4907a5112ec67e2d02a6
- Publication categories
- ; ;
- Author
- Pages
- [90]
- Substantive notes
- Sesja: Analytical Chemistry
- Miejsce wyd. i miejsce konf. wg siedziby org.
- Wydawca wg org. konf.
- Data wyd. wg daty konf.
- Book
- AMYC-BIOMED 2021 : 2nd Autumn Meeting for Young Chemists in Biomedical Sciences, 3-5 November 2021 – virtual conference : e-book of abstracts, 2021, [Naples], University of Naples Federico II, ISBN 978-88-94952-19-3
- Keywords in English
- fluorescent probe, BSA, albumin
- URL
- https://amyc-biomed-2021.webnode.it/chi-siamo/ Opening in a new tab
- Related project
- Diamentowy Grant "Projektowanie i synteza nowych sensorów luminescencyjnych oraz ich fotochemiczne badania potencjalnej przydatności do selektywnego i wielofunkcyjnego zastosowania w diagnostyce molekularnej. . Project leader at PK: , ,
- Language
- eng (en) English
- License
- Score (nominal)
- 0
- Uniform Resource Identifier
- https://cris.pk.edu.pl/info/article/CUT7f4859bc081a4907a5112ec67e2d02a6/
- URN
urn:pkr-prod:CUT7f4859bc081a4907a5112ec67e2d02a6
* presented citation count is obtained through Internet information analysis, and it is close to the number calculated by the Publish or PerishOpening in a new tab system.