Tan, Cheau Yuaan and Lim, Chun Shen and Liew, Siew Mun and Abd Halim, Adyani Azizah and Tayyab, Saad (2021) Lysine modification of human serum albumin and its effect on protein conformation and nalidixic acid binding. Journal of the Indian Chemical Society, 98 (3). ISSN 0019-4522, DOI https://doi.org/10.1016/j.jics.2021.100031.
Full text not available from this repository.Abstract
In order to investigate the involvement of lysine residues of human serum albumin (HSA) in nalidixic acid (NA) binding, various modified preparations of HSA such as 44% carbamylated (C-44), 83% carbamylated (C-83) and 85% acetylated (A(85)) were made by treating the HSA solution with a different molar excess of potassium cyanate and acetic anhydride. The extent of modification, charge homogeneity and conformational changes of these derivatives were checked by TNBSA reaction method, polyacrylamide gel electrophoresis (PAGE) and gel filtration using Sephacryl S-200 HR column, respectively. Binding of NA to HSA and its derivatives was examined using fluorescence quenching titration method to determine the binding constant. The emergence of a single band in PAGE and single symmetrical peak in gel filtration results confirmed the charge and size homogeneity of these derivatives. Hydrodynamic properties such as Stokes radius and frictional ratio, as obtained from the analytical gel filtration results suggested molecular expansion in C-83 and A(85) HSAs while C-44 HSA retained the native conformation. Addition of NA to both native and modified HSA derivatives quenched the fluorescence intensity of the protein at 344 nm to a different extent. Whereas the values of the Stern-Volmer constant (K-SV) and bimolecular quenching rate constant (k(q)) suggested, NA-HSA complex formation, binding constant (K-a) value suggested an intermediate binding affinity between NA and HSA. Furthermore, the decrease in the K-a value with the extent of modification was indicative of the involvement of lysine residues in NA-HSA interaction.
| Item Type: | Article |
|---|---|
| Funders: | Universiti Malaya (RG012/09AFR) |
| Uncontrolled Keywords: | Chemical modification; Human serum albumin; Drug-protein interaction; Nalidixic acid |
| Subjects: | Q Science > QD Chemistry R Medicine > RK Dentistry |
| Divisions: | Faculty of Dentistry |
| Depositing User: | Ms Zaharah Ramly |
| Date Deposited: | 01 Jul 2022 04:17 |
| Last Modified: | 01 Jul 2022 04:17 |
| URI: | http://eprints.um.edu.my/id/eprint/27994 |
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