Human serum albumin interaction with oxaliplatin studied by capillary isoelectric focusing with the whole column imaging detection and spectroscopic method
- 1 November 2009
- journal article
- research article
- Published by Elsevier BV in Journal of Pharmaceutical and Biomedical Analysis
- Vol. 50 (4), 570-575
- https://doi.org/10.1016/j.jpba.2008.10.028
Abstract
Capillary isoelectric focusing (CIEF) with whole column imaging detection (WCID) was used to investigate drug-protein interactions. This study was designed to examine the interaction between the platinum-based anticancer drug, oxaliplatin, with human serum albumin (HSA) in aqueous solution at physiological pH with drug concentrations of 10 to 100 μM and a constant concentration of HSA (5.0 × 10−5 M). The reaction mixtures were incubated for 0, 0.5, 1, 12, 24, 48 and 72 h at 37 °C in a water bath. The CIEF results indicate that with increasing the drug concentration, the complex formation of protein adducts increased compared to low-drug concentrations and major structural changes were observed as the incubation time progressed. The altered CIEF profile demonstrated the possible conformation change due to the binding of the drug. Results also showed a significant protein's pI shift for higher HSA–oxaliplatin incubation ratios. Furthermore, spectroscopic evidence shows that oxaliplatin caused the fluorescence quenching of HSA by formation of HSA–oxaliplatin complex. Using the Stern–Volmer equation, the quenching constants were calculated in the linear range. The quenching rate constants Kq at three different temperatures indicating the presence of static quenching mechanism in the interactions of oxaliplatin with HSA. This paper describes the validity of the CIEF-WCID technique for the study of protein–drug interactions and provides useful information and insight into the interaction of anticancer drugs with HSA.Keywords
This publication has 39 references indexed in Scilit:
- Dynamic Kinetic Capillary Isoelectric Focusing: A Powerful Tool for Studying Protein−DNA InteractionsAnalytical Chemistry, 2007
- Protein thermal stability and phospholipid–protein interaction investigated by capillary isoelectric focusing with whole column imaging detectionJournal of Separation Science, 2006
- Capillary Isoelectric Focusing Coupled with Dynamic Imaging Detection: A One-Dimensional Separation for Two-Dimensional Protein CharacterizationJournal of Proteome Research, 2006
- Characterization of phospholipid–protein interactions by capillary isoelectric focusing with whole-column imaging detectionAnalytical Biochemistry, 2006
- Role of calcium binding in protein structural changes and phospholipid–protein interactions studied by capillary isoelectric focusing with whole column imaging detectionAnalytica Chimica Acta, 2006
- Dynamic process of phospholipid–protein interaction studied by capillary isoelectric focusing with whole‐column imaging detectionElectrophoresis, 2006
- Characterization of bovine serum albumin–tryptophan interaction by capillary isoelectric focusing with whole column imaging detectionJournal of Chromatography A, 2006
- Capillary Isoelectric Focusing of Proteins with Liquid Core Waveguide Laser-Induced Fluorescence Whole Column Imaging DetectionAnalytical Chemistry, 2003
- Improved understanding in platinium antitumour chemistryChemical Communications, 1996
- Adaptation of the equipment for high-performance electrophoresis to isoelectric focusingJournal of Chromatography A, 1985