A simple method for obtaining human albumin and its use for in vitro interaction assays with indole-thiazole and indole-thiazolidinone derivatives
| Autor: | Luiz Bezerra de Carvalho Júnior, Sinara Mônica Vitalino de Almeida, Antônio Edson de Souza Lucena, Gleyton Leonel Silva Sousa, Josival Emanuel Ferreira Alves, Aurenice Arruda Dutra das Merces, Rafael David Souto de Azevedo, Maria Luiza Cavalcanti Lucena, Maria do Carmo Alves de Lima, Ricardo Olímpio de Moura |
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| Rok vydání: | 2021 |
| Předmět: |
Indoles
Serum Albumin Human Molecular Dynamics Simulation Biochemistry chemistry.chemical_compound Structural Biology Lowry protein assay medicine Blood plasma fractionation Humans Thiazole Molecular Biology Indole test Chromatography Binding Sites Molecular Structure Chemistry Tryptophan Albumin General Medicine Human serum albumin Fluorescence body regions Molecular Docking Simulation Thiazoles Spectrometry Fluorescence embryonic structures Thermodynamics medicine.drug Protein Binding |
| Zdroj: | International journal of biological macromolecules. 192 |
| ISSN: | 1879-0003 |
| Popis: | This work aimed to develop a simple and low-cost method to obtain human serum albumin (HSA) and its consequent application for in vitro drug interaction assays. The HSA was purified by classic principles of plasma precipitation and thermocoagulation, using a multiple-stage fractionation. The quality of the final product was assessed by electrophoresis, protein dosage by the Lowry method and the pharmacopeial thermal stability. At the end, an isotonic solution of HSA with a total protein concentration of 2.7 mg·mL−1 was obtained, which was visualized as a single band corresponding to the molecular weight of 66 kDa. After the thermal stability test, there was no indication of turbidity or color change of the solution. Finally, the HSA was useful for interaction assays with indole-thiazole and indole-thiazolidinone derivatives through UV–vis absorption and fluorescence spectroscopic studies, as well as by docking molecular analysis. Derivatives quenched the intrinsic fluorescence of HSA, disrupted the tryptophan residues microenvironment, and probably bind at Sudlow's site I. Therefore, the simplified methodology developed in this work proved to be effective in obtaining HSA that can be applied to research goals including drug interaction assays. |
| Databáze: | OpenAIRE |
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